TW201242598A - Combination therapies for hematologic malignancies - Google Patents

Abstract

The invention provides methods that relate to a novel therapeutic strategy for the treatment of hematological malignancies and inflammatory diseases. In particular, the method comprises administration of a compound of formula A, wherein R is H, halo, or C1-C6 alkyl; R' is C1-C6 alkyl; or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient; and one or more additional therapeutic agents optionally selected from the group consisting of bendamustine, rituximab, and ofatumumab.

Description

201242598 VI. Description of the invention: [Technical field to which the invention pertains] This application belongs to the fields of therapeutics and medicinal chemistry. In particular, this application relates to the use of certain salivation derivatives in combination with other therapeutic therapies for the treatment of hematological malignancies and certain other conditions. The application of the relevant application also cites the US Provisional Patent Application No. 61/452,03 4, filed on March 11, 2011, and the US Provisional Patent Application No. 61/ filed on June 3, 2011. 493,317, the priority of which is hereby incorporated by reference in its entirety. [Prior Art] Cellular signaling via 3' phosphorylated phosphoinositide has been implicated in a variety of cellular processes such as malignant transformation, growth factor signaling, inflammation, and immunity. The enzyme phospholipid inositol 3 kinase (PI 3K; PI3K), which produces these phosphorylated signaling products, initially identifies its activity in association with viral oncoproteins and growth factor receptor tyrosine kinases. Lipid creatinine (PI) and its phosphorylated derivatives are deacidified at the 3's base of the inositol ring. Xianxin PI 3 kinase activation involves a variety of cellular responses, including cell growth, differentiation, and apoptosis. Initial purification and molecular selection of PI 3 kinase revealed a heterodimer consisting of p85 and pi 10 subunits. Four different Class 1 PI3K's have been identified as ΡΙ3Κα, ΡΙ3Κβ, ΡΙ3Κδ, and ΡΙ3Κγ, each consisting of different 110 kDa catalytic subunits and regulatory subunits. More specifically, the three 163053.doc 201242598 subunits, that is, ρ110α, ρ110β, and ρΙΙΟδ each interact with the same regulatory subunit ρ85; and ρΙΙΟγ interacts with the different regulatory subunit ρ101. Each of these Κ3Κ has different expression patterns in human cells and tissues. • The identification of ρ 3 kinase ρ 110δ isoforms has been described in Chantry et al., / valence / C/zew, 272: 19236-41 (1997). Human V P11 〇δ isoforms were observed to be expressed in a tissue-defined manner. It is expressed in high amounts in lymphocytes and lymphoid tissues, suggesting that this protein may play a role in ΡΙ3 kinase-mediated signaling in the immune system. ΙΙΟ3Κ's ρΙΙΟβ isoforms also play a role in Κ3Κ-mediated signaling in certain cancers. There is a need for a therapy for a condition mediated by cancer, inflammatory diseases, and autoimmune diseases. In particular, there is a need for a therapy for cancers such as leukemias and lymphomas that are hematological malignancies. Existing chemical immunotherapy for chronic lymphocytic leukemia (CLL) can reduce the proliferation of malignant lymphocytes and enhance their apoptosis; however, because existing drugs may not effectively penetrate into these sites and because non-malignant stromal cells will be malignant lymphocytes Cells provide support, so these beneficial effects may be limited in lymph nodes. Existing chemotherapy or immunotherapy for CLL includes cyclophosphamide, chlorambucil, fludarabine, rituximab, alemtuzumab and Orfarizumab. These therapies are particularly effective in killing circulating malignant lymphocytes, but the effectiveness of reducing malignant lymphadenopathy may be insufficient. In addition, the treatments such as 163053.doc 201242598 do not induce lymphocyte redistribution or cause lymphocytosis. There is a need in the art for a therapy that treats CLL patients that modulates chemotaxis signaling between lymphocytes and stromal cells. In addition, there is a need for a therapy that reduces the effects of such signaling to redistribute malignant lymphocytes from the lymph nodes into the circulation. This redistribution may allow the chemotherapeutic or immunotherapeutic agent to better kill such cells. SUMMARY OF THE INVENTION The present application satisfies the needs of the art by providing a method of using such compounds in combination with other chemotherapeutics or immunotherapies to treat cancer, inflammatory diseases, and autoimmune diseases. Specifically, cancers, such as leukemias and lymphomas, are treated as hematological malignancies using the methods described herein. In one aspect, a method of treating cancer comprising administering an effective amount of a compound of formula A to an individual in need of such treatment,

HN丫, (Formula A), n^Yn

Wherein R is hydrazine, a aryl group or a C1-C6 alkyl group; |^C1_C6 alkyl; or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient; A variety of other therapeutic agents selected from the group consisting of bendamustine, rituximab, and orfarizumab are optionally selected. 163053.doc 6 - 201242598 In one embodiment, the compound is

In another embodiment, the compound is

In some of the above embodiments, the cancer is a hematological malignancy. In some of the above embodiments, the hematological malignancy is a B cell malignant tumor. In some of the above embodiments, the hematological malignancy is leukemia or lymphoma. In some of the above embodiments, 'the cancer is chronic lymphocytic leukemia (CLL) or non-Hodgkin's lymphoma (n〇n H〇dgkin, s, NHL). In some embodiments, the cancer is inert non-Hodge Gold lymphoma (iNHL) 其他 In other embodiments, each compound and one or more therapeutic agents are administered at least once during each cycle, and one or more of the therapeutic agents are 163053.doc 201242598 in and with Individuals are administered to the same or different cycles. In some of the above embodiments, the period is 7 to 42 days. In some of the above embodiments, the _ or therapeutic agents are administered to the individual on at least the first and second days of at least one cycle. In some of the above embodiments, the one or more therapeutic agents are administered to the individual weekly for at least one period. In some of the above embodiments, the compound is administered to the individual between 5 and 2 mg twice a day. In some of the above embodiments, the individual is administered one or more therapeutic agents between 50 mg/m2 and 1,500 mg/m2. In some of the above embodiments, the compound is present in a pharmaceutical composition comprising a compound and at least one pharmaceutically acceptable excipient. In some of the above embodiments, the individual is resistant to standard chemotherapeutic therapies. In some of the above embodiments, the individual has at least one enlarged lymph node. In some of the above embodiments, the individual i} is refractory to at least one type of chemotherapy treatment, or Π) relapses after chemotherapy treatment, or a combination thereof. In another aspect, the invention provides a method of treating cancer comprising administering to a subject in need of such treatment an effective amount of a compound of formula A,

And pharmaceutically acceptable salts thereof; And one or more other therapeutic agents selected from the group consisting of bendamustine, rituximab, orfarizumab, and lenalidomide, as appropriate. In another aspect, a method of treating a condition comprising administering to an individual in need of such treatment an effective amount of a formula "

(II") or a pharmaceutically acceptable salt thereof, and or a plurality of other therapeutic agents selected from the group consisting of: bentazol; rituximab and orfarizumab, Cellular leukemia (CLL) or inert non-hoof, wherein the condition is chronic lymphduritis (iNHL). The treatment of chemotherapy is difficult or a combination thereof. In one embodiment, the individual i) is at least one cured , or ii) recurring after chemotherapy treatment, in the above-mentioned embodiments, the 'compound and/or the plurality of therapeutic agents are each administered at least one, during the period of one week, and at least one person, and wherein - or A plurality of treatments are administered to the individual within the same or different periods of administration of the compound. In the above embodiments, the period is from 7 to 42 days. In the above embodiment, 50 mg is administered to the individual twice a day. 200 163053.doc 201242598 mg of the compound. In the implementation of a - or a variety of therapeutic materials, the above-mentioned embodiments, at least the first and second days of at least one cycle to the individual Injecting bendamose juice. In some of the above embodiments Investigate /, Bendamustine for at least 6 cycles. In some of the above examples, Benzene 2, 2t, lcn 2 Benzaloxidin Ding is dosed between 50 mg / m and 150 mg / Between m. In some of the above embodiments, the K method additionally comprises administering to the individual rituximab. In some of the above embodiments, at least the first day of each cycle of the community's 6 cycles Rituximab. < In another embodiment... or a variety of therapeutic rituximab. In some of the above-mentioned implementations, rituximab is administered weekly to individuals. In some of the above embodiments, each dose of rituximab-resistant is between 3. In another embodiment, or a plurality of therapeutic agents are olfaximab. 6 cycles of administration of at least (4) Orphea in another aspect, providing a method for the individual to be treated with an effective treatment of the condition, including the requirement of the formula I" or formula II" compound, 163053 .doc 201242598

Or a pharmaceutically acceptable salt thereof, and one or more other therapeutic agents selected from the group consisting of bendamustine, rituximab, orfarizumab and lenalidomide, as appropriate. In one embodiment, the one or more therapeutic agents are lenalidomide. In another aspect, a method of treating an individual having a B cell disorder, comprising: a) identifying an individual having a B cell malignancy, wherein the individual is refractory or at least at least one or more of the therapies Recurrence after treatment with one or more therapies selected from the group consisting of bortezomib (Velcade®), carfiizomib (PR-171), PR-047, disulfide ( Disulfiram), racttastatin (lactacystin), PS-519 'eponneniycin, epoxomycin, aclatinoniycin, CEP-1612, MG-132 'CVT-63417, PS-341, ethylene quinone tripeptide inhibitor, ritonavir, PI-083, (+/-)-7-mercapto OMrad ((+/-)-7 -methylomuralide), (-)-7-methyl omrad, perifosine, rituximab, sildenafil citrate (Viagra®), CC-5103, Thalidomide, erratuzumab (hLL2-anti-CD22 humanized antibody), xin 163053.doc 201242598 vavastatin Enzavirin (enzastaurin), Campath 1H®, dexamethasone, DT PACE, oblimersen, antinepraplast A10, anti-Xinpratong AS2 1 Ranibizumab, beta alexin, cyclophosphamide, doxorubicin hydrochloride, pegylated liposomal cranberry hydrochloride, prednisone, prednisolone ), cladribine, vincristine sulfate, fludarabine, filgrastim, melphalan, recombinant interferon a, carmustine ), cisplatin, cycloheximide, cytarabine, etoposide, mefaxine, dolastatin 10, indium In 111 monoclonal antibody MN-14 , 钇Y 90 humanized erarazumab, anti-thymocyte globulin, busulfan, cyclosporine, methotrexate 'mycophenolic acid' Mycophenolate mofetil, therapeutic allogeneic lymphocytes Y 90 Yttrium Y 90 ibritumomab tiuxetan, sirolimus, tacrolimus, carboplatin, thiotepa, paclitaxel Paclitaxel), aldesleukin, recombinant interferon alpha, docetaxel, ifosfamide, mesna, recombinant interleukin-12 ), recombinant interleukin-11, Bcl-2 family protein inhibitor ABT-263, denileukin diftitox, tanespimycin, everolimus, polyethyl Glycolation 163053.doc 12 201242598 Pegfilgrastim, vorinostat, avocifi, recombinant flt3 ligand, recombinant human thrombopoietin, lymphatic hormone (lymph) 〇kine) activation of killer cells, amifostine trihydrate, aminocamptothecin, irinotecan hydrochloride, caspofungin acetate (caSp0fungin ace) Tate), clofarabine, epoetin alfa, nelarabine, pentostatin, sargramostim, vinorelbine hydrogenate (vinoreibine) Ditartrate), WT-1 analog peptide vaccine, WT1 126-134 peptide vaccine, fenretinide, ixabepilone, oxaliplatin, monoclonal antibody CD19, monoclonal antibody CD20, omega-3 fatty acids, mitoxantrone hydrochloride, octreotide acetate, tositumomab, and moths 131 tosimozumab, motosone (motexafin gad〇linium), arsenic trioxide, tipifarrfib, autologous human tumor-derived HSPPC-96, veltuzumab, bryostatin 1), anti-CD20 monoclonal antibody, phenoxybutyrate, pentostatin, lumiliximab, apolizumab, anti-CD40, orfarizumab, Tianxi Temsirolimus, bendamus $, 嘌An analog, lenalidomide, an alkylating agent, and anthracycline-containing therapy or a combination thereof; b) administering to a subject in need of such treatment a compound I"compound 163053.doc (I"), (I "), 201242598

Or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient selected according to the conditions of the sun; and or more optionally selected from the group consisting of benzalkonium; butyl, rituximab and amine Other therapeutic agents of the group consisting of wood monoclonal antibodies. In one embodiment, the 'individual is refractory to at least one or more therapies or relapses after treatment with at least one or more therapies selected from the group consisting of rituximab, a home base, fluoride Dalabin and bismuth-containing ring-gel therapy and combinations thereof. In some of the above embodiments, between 50 mg and 200 mg of the compound is administered to the individual twice daily during at least one or more cycles. In some of the above embodiments, one or more additional therapeutic agents between 50 1 ^/1112 and 1,500 mg/m 2 are administered to the individual at least once during at least one or more cycles of the one or more therapeutic agents. The individual is administered within the same or different period as the administered compound. In another aspect, a method of treating an individual having a B cell disorder, comprising: a) identifying an individual having a B cell malignancy, wherein the individual is refractory or at least at least one or more of the therapies Recurrence after treatment with one or more therapies' Therapies are selected from the group consisting of bortezomib 163053.doc •14-201242598 (Velcade®), Kafizomi (PR-171), PR-047, disulfiram , ractaxietine, PS-519, ebonymycin, epoxymycin, aclarithromycin 'CEP-1612, MG-132, CVT-63417, PS-341, vinyl sulfone tripeptide inhibitor, Ritonavir, PI-〇83, (+/_)_7_methyl OMrad, Bu)·' 曱基奥姆拉德'Perifusone, rituximab, sildenafil lemon Acidate (Viagra®), CC-5103, Thalidomide, Irlatuzumab (hLL2-anti-CD22 Humanized Antibody), Simvastatin, Enzatoline, Campath 1H®, Dexamethasone, DT PACE, Olimpson, anti-New Praton A1 0, anti-nepraplatin AS2 1, aleizumab, beta alixine, cyclosporin, cranberry hydrochloride 'PEGylation Small liposome Raspberry hydrochloride, prednisone, prednisolone, cladribine, vincristine sulfate, fludarabine, filgrastim, melphalan, recombinant interferon alpha, carmustine, cisplatin, ring Phosphonamine, cytarabine, etoposide, melphalan, dolphin toxin 1 铟, indium In 111 monoclonal antibody ΜΝ-14, 钇Υ 90 humanized erarazumab anti-thymocyte globulin , busulfan, cyclosporine, methotrexate, mycophenolate, therapeutic allogeneic lymphocytes, y Y 90 tiltaneximumab, sirolimus, tacrolimus, card Platinum, thiotepa, paclitaxel, adiponectin, recombinant interferon alpha, paclitaxel, ifosfamide, mesna, recombinant interleukin-12, recombinant interleukin -11, Bcl-2 Family protein inhibitor ABT-263, Dinixin fusion toxin, Tansone, Everolimus, PEGylated filgrastim, vorinostat, avoxime, recombinant flt3 ligand , recombinant human thrombopoietin, lymphocyte-activated killer cells, amine phosphinine trihydrate, amine camptothecin, irinotecan hydrochloride, card Pofin acetate, cloprobine, afar berberine, naira 163053.doc -15· 201242598 pr, pentastatin, sagstatin, vinorelbine tartrate, chlorin 丨 丨 analog peptide Vaccine, WT1 126_134 peptide vaccine, retinoic acid, ixabepilone, acesulfide platinum, monoclonal antibody CD19, monoclonal antibody CD2 〇, ω 3 fatty acid, mitoxantrone hydrochloride, octreotide acetate, Torr Simuzumab and iodine 131 tosimozumab, motosone, arsenic trioxide, teprefinib, autologous human tumor-derived HSPPC_96 'Ventuzumab, bryostatin, anti-CD20 single Antibody, chlorambucil, pentastatin, rufuzumab 'aborizumab, anti-CD40, orfarizumab, sirolimus, bendamustine, guanidine analogue, come Nalamine, alkylating agent and anthracycline-containing therapy or a combination thereof; b) administering a compound of the formula to an individual in need of such treatment

Or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient; and/or a plurality of optionally selected from the group consisting of bendamus; rituximab, olfaximab Other therapeutic agents that are resistant to the group of lenalidomide. In another aspect, the invention provides a method of treating chronic lymphocytic leukemia (CLL) or indolent non-Hodgkin's lymphoma (iNHL) comprising administering to an individual in need of such treatment an effective amount of formula j&quot ; compound, 163053.doc -16- 201242598

Or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable excipient selected as appropriate; and bendamustine. The individual compound and the bendamus lamp are administered at least once a week for at least one week, and wherein the bendamust; τ line is administered to the individual within the same or different period as the administered lysate. In some of the above embodiments, the period is 7 to 42 days. In some of the above embodiments, the 'compound is administered to the individual twice every at least one cycle' and wherein the bendam 5 cleft is administered to the individual for at least the first and second days of at least one cycle. In some of the above embodiments, the bendamustine is administered for at least 6 cycles. In some of the above embodiments, the dose of the compound is between 50 and 200, and wherein the dosage of stupa is 50 mg/m2 and 15 mg/m2. In some of the above embodiments, the method further comprises administering one or more additional therapeutic agents selected from the group consisting of rituximab and orfarizumab. In some of the above embodiments, the method further comprises administering - or a plurality of other treatments selected from the group consisting of rituximab anti-olfaximab and lenalidomide I63053.doc • 17· 201242598 in another In one aspect, a method of treating chronic lymphocytic leukemia (CLL) or inert non-Hodgkin's lymphoma (iNHL) comprising administering an effective amount of a compound to a subject in need of such treatment,

Or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable excipient selected as appropriate; and rituximab. In one embodiment, the compound and rituximab are each administered at least once in the cycle and wherein the rituximab is administered to the individual within the same or different period of time as the administered compound. In some of the above embodiments, the period is 7 to 42 days. In the above-described implementation, the compound is administered to the individual twice a day during at least one cycle, and wherein the rituximab is administered to the individual weekly for at least one cycle. In some of the above embodiments, the dose of the compound is between 50 mg and 200 mg' and wherein the dose of rituximab is between _mg/m2 and mg/m2. The process of the above-described embodiments additionally comprises administering one or more additional therapeutic agents selected from the group consisting of bendamus>, butyl and olfaxim, and imazonb. In some of the above embodiments, the method additionally comprises administering one or more other therapeutic agents selected from the group consisting of benzodiazepine/T, orfarizumab and lenalidomide selected from 163053.doc 201242598. In another aspect, a method of treating chronic lymphocytic leukemia (called or inactive non-Hodgkin's lymphoma (iNHL) comprising administering an effective amount of a compound to a subject in need of such treatment is provided,

Or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable excipient selected as appropriate; and lenalidomide. In one embodiment, the compound and lenalidomide are each administered at least once during the cycle, and wherein the lenalidomide is administered to the subject within the same or different period of time as the administered compound. In another aspect, a method of treating chronic lymphocytic leukemia (CLL) or inert non-Hodgkin's lymphoma (iNHL) comprising administering an effective amount of a compound of the formula to an individual in need of such treatment,

163053.doc 201242598 or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable excipient selected as appropriate; and olfaximab. In some of the above examples, the dose of the compound was between 5 mg and 2 mg, and the dose of orfarizumab was between 200 mg and 1500 mg. In some of the above embodiments, the initial dose administered to olfaxumab is different from the subsequent dose of olfaxumab. [Embodiment] All technical terms, notations, and other scientific terms or terms used herein are intended to have a meaning commonly understood by those skilled in the art to which the invention pertains, unless otherwise claimed. In some instances, terms that have a commonly understood meaning are defined herein for the sake of clarity and/or for ease of reference, and the inclusion of such definitions herein is not necessarily construed as indicating that it is generally understood in the art. There are substantial differences in the meaning. Many of the techniques and procedures described or referenced herein are well understood by those of ordinary skill in the art and are generally employed Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed according to the manufacturer's protocol and/or parameters, unless otherwise stated. The discussion of the general methods given herein is intended for illustrative purposes only. Other alternative methods and embodiments will become apparent to those skilled in the art from reviewing this disclosure. A group of items linked to the conjunction "or" shall not be deemed to require mutual exclusivity in the group, but shall be "and/or" unless otherwise stated. Although the items, elements, or stipulations of the present invention may be described or claimed in the singular, the invention is intended to be The present invention provides methods for novel therapeutic strategies for treating cancer and inflammatory diseases. In one aspect, the invention provides a method of treating cancer in an individual comprising administering to the individual a compound of formula A

Wherein R is H, halo or C1-C6 alkyl; R. is C1-C6 alkyl; or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient. In a particular embodiment, the halo group is F; and R is methyl, ethyl or propyl. In a particular embodiment, 'R is attached to the 5 position of the quinazoline ring, which has a structure

In a particular embodiment, R is attached to the 6 position of the quinazolinyl ring, the ring having the structure 163053.doc -21 · 201242598

The term "compound" as used herein, unless otherwise specified, refers to a gas A compound such as Compound I, Compound π, or an enantiomer (such as I" or II"), or a mixture of enantiomers. "Compound of Formula I" or "Compound I" means a compound 5-fluoro-3-phenyl [1-(9Η·嘌呤-6-ylamino)-propyl]_3H-quinazoline-4-_, having the formula ^士建:, '.

The S-enantiomer of Compound I, shown as hydrazine, is shown here:

"Formula π compound" or "compound „" means a compound 嘌呤-6-ylamino)ethyl)-6-fluoro-3-phenyl quinazoline _4(3Η)-one having the structure of formula II: 163053.doc •22- 201242598

The s-enantiomer of Compound II is shown here and is called Η":

In one embodiment, the hydrazine compound is a compound of formula I. In another embodiment, the compound of formula A is a compound of formula π. In certain embodiments, the compound is a racemic mixture of the R-enantiomer and the S-enantiomer. In certain embodiments, the compounds are employed as a mixture of enantiomers, and are typically enriched in the S-enantiomer. In some embodiments, the compound is predominantly the S-enantiomer. In some embodiments, the compound of formula A used in the methods described herein is at least 80% S-enantiomer. In certain embodiments the compound consists essentially of the S-enantiomer, wherein the compound comprises at least 66-95% or 85-99% of the S-enantiomer. In some embodiments, the compound has an s-enantiomer of at least 90% or at least 95% enantiomeric excess (e e ). In some embodiments, the compound has at least 98% or at least 99. /. S-enantiomeric excess (in some embodiments, the compound comprises at least 95% of the S. Enantiomer 4 examples provided by: 163053.doc • 23· 201242598 Cell and Patient Experiments In the particular example, the sample of the compound used is more than 95% s enantiomer. In a particular embodiment, the compound of formula ι or formula n used in the methods described herein is at least 80% S-enantiomer. In certain embodiments, the compound of formula or formula π consists essentially of the S-enantiomer, wherein the compound comprises at least 66-95% or 85-99% of the 8 enantiomer. In some embodiments The compound of formula II has an enantiomeric excess of at least 90% or at least 95% (the S-enantiomer of .... In some embodiments, the compound of formula ! or formula π has at least 98% or at least 99% S-enantiomeric excess (ee). In certain embodiments, the 'U or formula (10) compound comprises at least 95% of the 8-enantiomer. In the cell and patient experiments provided in the Examples section The sample of the compound I used is 95 ° /. The above S-enantiomer. In a specific implementation, compared to other PI3K isoforms, compound Selective inhibition of ΡΙ3Κ ρ 110δ. In a particular embodiment, the cancer is a hematological malignancy and/or a solid tumor. In another- (four) embodiment, the 'hematological malignancy is leukemia or lymph. In some embodiments, the lymphoma is Mature (peripheral cutaneous cell tumors. In a particular embodiment, the mature sputum cell tumor is selected from the group consisting of: sputum cells chronic lymphocytic leukemia/small lymphocytic lymphoma; sputum cells Prolymphocytic ieukemia; LymPh〇plasmacytic lymphoma; marginal zone lymphoma (10) 咕w zone lymphoma), such as spleen marginal zone B-cell lymphoma (+/velillary lymph 163053. Doc •24- 201242598 cells), nodular marginal zone lymphoma (+/- monocyte-like B cells) and mucosa-associated lymphoid tissue (MALT) type nodular peripheral zone B-cell lymphoma; Hairy cell leukemia ; polycytokines/plasmacytoma; Follicular lymphoma (follicle center); mantle cell lymphoma (Mantle c Ell lymphoma); diffuse large cell B-cell lymphoma (including mediastinal large B-cell lymphoma, intravascular large B-cell lymphoma, and primary effusion lymphoma) And Burkitt's lymphoma/Burkitt's cell leukemia. In some embodiments, the lymphoma is selected from the group consisting of multiple myeloma (MM) and non-Hodgkin's lymphoma (NHL), mantle cell lymphoma (MCL), bubial lymphoma, val Waldenstrom's macroglobulinemia (WM) or B-cell lymphoma and diffuse large B-cell lymphoma (DLBCL). In another specific embodiment, the leukemia is selected from the group consisting of acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and small lymphocytic lymphoma (SLL) ). Acute lymphocytic leukemia is also known as acute lymphoblastic leukemia and is used interchangeably herein. Both terms describe a type of cancer that begins with white blood cells in the bone marrow, i.e., lymphocytes. In some embodiments, non-Hodgkin's lymphoma (NHL) belongs to one of two species, invasive NHL or inert NHL. Invasive NHL grows rapidly and can lead to relatively rapid death in patients. Untreated survival can be several months or 163053.doc -25· 201242598 to weeks. Examples of invasive NHL include B cell tumors, diffuse large B cell lymphoma, T/NK cell tumors, anaplastic large cell lymphoma, peripheral T cell lymphoma, prodromal lymphoma Cell leukemia/lymphoma, precursor T lymphoblastic leukemia/lymphoma, Burkitt's lymphoma, adult T-cell lymphoma/leukemia (HTLV1+), primary CNS lymphoma, mantle cell lymphoma, polymorphism Post-transplant lymphoproliferative disorder (PTLD), AIDS-associated lymphoma, true histiocytic lymphoma, and parental nk cell lymphoma. The most common type of invasive NHL is diffuse large cell lymphoma. Inert NHL grows slowly and does not show significant symptoms for most patients until the disease has progressed to advanced stages. The untreated survival of patients with indolent NHL can be years old. Non-limiting examples include follicular lymphoma, small lymphocytic lymphoma, marginal zone lymphoma (such as extranodal marginal zone lymphoma (also known as mucosa-associated lymphoid tissue-MALT lymphoma), nodular marginal zone b-cells Lymphoma (monocyte-like B-cell lymphoma), spleen marginal lymphoma) and lymphoplasmacytic lymphoma (Waldenstrom's macroglobulinemia). In some cases, tissue transformation can occur, such as the patient's inertia, which can be converted to invasive NHL. In some implementations, the invention provides a method of treating a patient suffering from invasive nhl or indolent NHL. In some embodiments, the invention provides methods of treating a patient having a condition selected from the group consisting of: mantle cell lymphoma (mcl), diffuse large B-cell lymphoma (DLBCL), buoyancy Lymphoma _, acute lymph 163053.doc •26- 201242598 Cellular leukemia (ALL), acute bone marrow leukemia (aml), chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (sll), multiple bone marrow Tumor (MM) and marginal zone lymphoma. In some embodiments, the methods of the invention are administered to a patient having a relapsed or refractory condition. In another example, the cancer is breast cancer, lung cancer, colon cancer or prostate cancer. In a specific embodiment, the cancer is associated with abnormal Κ3Κ activity compared to ρΐ3κ activity in a cancer-free individual. In a particular embodiment, the preferred individual is difficult to administer (4) or relapses after chemotherapy treatment. In an alternative embodiment, the individual is a primary (de 77〇V〇) patient. In a particular embodiment, the method comprises reducing the degree of p-activity in the patient. In a particular embodiment, the individual is a human subject. Individuals undergoing treatment with known therapeutic agents may experience resistance to treatment. For example, although FDA approved frotezomib for relapsed/refractory, relapsed, and newly diagnosed MM's, some patients did not respond to bortezomib and other patients acquired resistance to ezastigmine. In some embodiments, the linalone compounds described herein synergistically enhance the efficacy of known therapeutic agents. In some embodiments, the compounds described herein can enhance any of the therapeutic agents described herein. In a more specific embodiment, the compounds described herein synergistically enhance a proteasome (Pr〇tea S〇me) inhibitor. In some of the above embodiments, the individual is resistant to chemotherapeutic therapies. In some of the above embodiments, the individual is resistant to the protein 163053.doc -27-201242598 clonal inhibitor. In some of the above embodiments, the individual is resistant to stilbenzol or carbomizomib. In one example, the compounds described herein synergistically enhance the detoxification induced by bortezomib. Without being bound by theory, the compounds discussed herein inhibit ketotizil-induced AKT disc acidification. In some embodiments, the methods described herein are used to overcome resistance to proteasome inhibitor therapy. In some embodiments, the invention provides methods of treating an individual who is resistant to, or has developed resistance to, a therapeutic agent. While not being bound by theory, the synergistic effect between a compound of formula A and conventional therapies can be attributed to the ability of the compounds of the invention to induce tumor cells to move into the peripheral circulation. The peripheral circulation of sputum tumor cells enhances the ability of conventional therapies to act and more effectively neutralize tumors. This synergy has been demonstrated in patients. Thus, the method comprises administering to the patient, in addition to the compound of formula A, at least a therapeutically effective amount of at least one additional therapeutic agent and/or treatment route selected to treat the cancer in the patient. "Therapeutic agent" may mean one or more compounds as used herein. The therapeutic agent can be a standard or experimental chemotherapeutic drug. The therapeutic agent can comprise a combination of more than one chemotherapeutic agent. A typical chemotherapeutic combination of drugs is a_q^ listed herein, and a particular therapeutic agent can be selected depending on the type of disease being treated. Non-limiting examples of conventional chemotherapeutic therapies for specific blood disorders are described in subsequent sections. In a particular embodiment, the invention provides a method of treating a hematopoietic cancer patient, such as a CLL patient, with bortezomib and a compound of formula A (e.g., Formula 1, oxime, 1 or guanidine), wherein the combination provides a synergistic effect. .doc • 28 - 201242598 In a particular embodiment, the therapeutic agent is selected from the group consisting of: bortezomib (Velcade®), kafizomib (PR_171), pR 〇47, disulfiram, thunder Ketaxi 'Ting, PS-519, Ebonymycin, epoxymycin, aclarithromycin, CEP-1612, MG-132, CVT-63417, PS-341, vinyl sulfone tripeptide inhibitor, benefit Tonavir, PLOM, (+/_)_7•曱基奥姆拉德, (-)-7-曱基奥姆rad, 派瑞福松, rituximab, sildenafil citrate (Viagra®), CC-5103, Thalidomide, Irlatuzumab (hLL2-anti-CD22 Humanized Antibody), Simvastatin, Enda Tallin, Campath-ΙΗ®, Dexamethasone, DT PACE, Olimpson, anti-New Praton A10, anti-nepraplatin AS2-1, aleuzumab, beta alexin, cyclosporin, cranberry hydrochloride, PEGylation fat Plastid red per hydrochloride, prednisone, prednisolone, cladribine, vincristine sulfate, lactapamil, filgrastim, methadine, recombinant interferon alpha, carmustine, Cisplatin, cyclophosphamide, cytarabine, etoposide, melphalan, dolastatin 10, indium In 111 monoclonal antibody ΜΝ-14, 钇Υ90 humanized erarazumab, anti-thymus Cytoglobulin, busulfan, cyclosporine, methotrexate, mycophenolate mofetil, therapeutic allogeneic lymphocytes, 钇γ 9 〇 坦 异 bembezumab, sirolimus, he Chlorhexidine, carboplatin, thiotepa, paclitaxel, adiponectin, recombinant interferon alpha, paclitaxel, ifosfamide, mesna, recombinant interleukin-12, recombinant interleukin 11. Bcl-2 family protein inhibitor ΑΒΤ-263, lignin fusion toxin, tan spiromycin, everolimus, pegylated filgrastim, vorinostat, avoxe, Recombinant flt3 ligand, recombinant human thrombopoietin, lymphokine-activated killer cell, amine phosphine trihydrate, amine-based hi tree 163053.doc -29· 2 01242598 Alkaline irinotecan hydrochloride, caspofungin acetate, clolapabine, afaribine, statin, naribine, pentastatin, sagstatin, vinorelbine tartrate, state analog Peptide vaccine, WT1 126_m peptide vaccine, retinoic acid, esapanidine H, serotonin cdi9, monoclonal antibody CD20, ω_3 fatty acid, mitoxantrone hydrochloride, octreotide acetate 'tosimo Monoclonal antibody and iodine! 131 tosimozumab, motosone, arsenic trioxide, teprefinib, autologous human tumor-derived Hsppc_96, virulzumab, tyrosin 1, anti-CD20 Antibody, chlorambucil, pentastatin, luciximab, apocilizumab, anti-(:1)4〇 and orfarizumab or a combination thereof. Combinations of therapeutic agents are used in current and experimental therapies, such as those combinations a-q listed above. In some embodiments, the therapeutic agent is preferably a proteasome inhibitor. In some embodiments, the methods comprise administering a compound and a proteasome inhibitor. Proteasome inhibitors include natural and synthetic compounds. Non-limiting examples of proteasome inhibitors include bortezomib ([(1R)_3_methylphenylpyrazine-2-ylcarbonyl)amino]propenyl}amino)butyl]-acid), which Marketed by Millennium Pharmaceuticals as "Velcade®"; Kafizomi (PR-171) and oral analogue pR-〇47, both developed by proteolix, Inc. Other examples of proteasome inhibitors include disulfiram; rektastatin; synthetic compounds such as PS-5 19, econazole, epoxymycin, and aclarithromycin; calpain inhibitors , such as CEP-1612, MG-132, CVT-63417, PS-341; ethylene quinone tripeptide inhibitor; ritonavir; PI-083; (+/-)-7-methyl OMrad; (-)-7-methyl OMrad. In a particular embodiment, the compound of formula A is administered in combination with bortezol 163053.doc • 30·201242598 meters or kafizomib. In a more specific embodiment, the compound of formula ι is administered in combination with bortezomib or carfilzomib. In other specific embodiments, the compound of formula π is administered in combination with bortezomib or carfilzomib. In a specific embodiment, the compound of formula A is administered in combination with rituximab or orfarizumab. In a more specific embodiment, the guanidine compound is administered in combination with rituximab or orfarizumab. In other specific embodiments, the π compound is administered in combination with rituximab or orfarizumab. In some embodiments, the therapeutic agent is an alkylating agent. Non-limiting examples of alkylating agents include, for example, busulfan, melphalan, chlorambucil, cyclophosphamide, mechlorethamine, uramustine, ifosfamide, Carmustine, lomustine (1〇mustine), streptozocin, thiotepa and platinum-based chemotherapeutic drugs such as cisplatin, carboplatin, nedaplatin, orsay Platinum, satraplatin, triplatin tetranitrate. In other embodiments, any of the compounds of the invention may be combined with one or more other active therapeutic agents in unit dosage form for simultaneous or sequential administration to a patient. Combination therapies can be administered simultaneously or sequentially. When administered sequentially, the combination can be administered by administering two or more administrations. In one embodiment, 'co-administering a compound of the invention and one or more additional/tongue therapeutic agents generally means administering the compound of the invention and one or more additional active therapeutic agents simultaneously or sequentially to achieve a therapeutically effective amount. The inventive compound and one or more other active therapeutic agents are present in the patient. In an alternative embodiment, the compound and therapeutic agent need not necessarily be present in the patient, but the specific time course of administration of the compound and therapeutic agent will produce a synergistic effect. 163053.doc • 31 · 201242598 co-administration includes administering a unit dose of a compound of the invention at or after administration of one or more other active therapeutic agents; for example, a few seconds after administration of one or more other active therapeutic agents, The compounds of the invention are administered in minutes, hours or days. For example, a unit dose of a compound of the invention may be administered first, followed by administration of one or more additional active therapeutic agents in a unit of time, minutes, hours, or days. Alternatively, one or more additional therapeutic agents can be administered first, followed by administration of a unit dose of a compound of the invention in seconds, minutes, hours or days. In some cases, it may be desirable to first administer a unit dose of a compound of the invention, followed by administration of one or more of the other active therapeutic agents after a period of hours (e.g., 1-12 hours). In other instances, it may be desirable to first administer one or more of the other active therapeutic agents, followed by administration of a unit dose of a compound of the invention after a period of hours (e.g., 1-12 hours). In some cases it may be desirable to first administer a unit dose of a compound of the invention, followed by administration of one or more additional active therapeutic agents in a unit dose over a period of days (e.g., 1-12 days). In other instances, it may be desirable to first administer one or more of the other active therapeutic agents, followed by administration of a unit dose of a compound of the invention after a period of days (e.g., 1-12 days). In some embodiments, the dosing regimen may involve alternating administration of the compound and therapeutic agent over a period of days, weeks, or months. Combination therapy can provide "synergistic effect" and "synergistic effect", that is, the effect achieved when the active ingredient is used together is greater than the sum of the effects produced by the compound alone. When the active ingredient: (丨) is co-formulated in the form of a combined formulation And at the same time, it is delivered or delivered; (2) alternately or in parallel in the form of separate formulations, 163053.doc • 32- 201242598; or (3) synergistic effects can be achieved according to a certain other scheme. When administered in alternation therapy, the compounds are administered or delivered sequentially, for example, in the form of separate troches, pills or capsules or by different injections of sputum injection 11: synergistic effects can be achieved in general, t, in alternation therapy During the period, the active ingredients in the active agent are administered sequentially (i.e., continuously). In one embodiment, the invention provides a pharmaceutical composition, a compound I, , a formula of 匕3 or a pharmaceutically acceptable excipient thereof. In one aspect, another compound:

Acceptable salts and at least one of the pharmaceutically acceptable compositions are enriched in the S-enantiomer. The invention provides a pharmaceutical composition, comprising the formula

Or a pharmaceutically acceptable salt thereof; and at least one excipient for treating hurricane. In a practical, acceptable embodiment, the composition is enriched in the S-enantiomer. 163053.doc -33- 201242598 In one aspect the invention provides a method of treating a multiple myeloma (MM) in a patient comprising administering a compound of formula A in combination with another therapeutic agent. In some embodiments 'Formula A is Compound (4). In a particular embodiment, 'Formula A is a compound worker. In other embodiments, Formula VIII is a compound Π". In some of the above embodiments, the other therapeutic agent is a proteasome inhibitor. In a particular embodiment, the additional therapeutic agent is bortezomib. In a particular embodiment, a method of treating multiple myeloma in a patient comprises administering Compound I" and frotezomib. In a particular embodiment, a method of treating a patient with multiple myeloma comprises administering a compound „” and frotezomib. In some of the above embodiments 'compound! "or Π" having an enantiomeric excess of at least 6 (%). In some of the above examples, the compound I" or Π" has an enantiomeric excess of at least 70%. In some of the above examples, The compound "" or π" has an enantiomeric excess of at least 8%. In some of the above examples, the compound I" or hydrazine has an enantiomeric excess of at least 90%. In some of the above embodiments, the compound j" or π" has an enantiomeric excess of at least %. In some of the above embodiments, the compound 〖" or π" has an enantiomeric excess of at least 98%. In some of the above embodiments, the compounding, or π " has an enantiomeric excess of at least 99%. In a particular embodiment, the combination of therapeutic agents is administered in combination with the compound, wherein The combination is selected from the group consisting of: a) bendamustine; b) rituximab; c) bendamustine and rituximab; d) olfaximab; and 163053. Doc • 34. 201242598 e) Lenalidomide. In an alternative embodiment, the compound is used in combination with a therapeutic procedure. In a particular embodiment, the therapeutic procedure is selected from the group consisting of: peripheral blood dry fine , self-destructive hematopoietic stem cell transplantation, autologous bone transplantation, antibody therapy, biological therapy, enzyme inhibitor therapy, total body irradiation, stem cell rounds, clearing bone marrow with stem cell support (b_marrow ablation whh (10) _ Μ—(1) living body Peripheral blood stem cell transplantation, umbilical cord blood transplantation, immunoenzymatic technique, immunohistochemical staining, pharmacological research, low let-drill, 6 〇 γ-ray therapy, ble〇mycin, conventional surgery, radiation Therapy, high-dose chemotherapy, and non-(four) clearance allogeneic (four) hematopoietic stem cell transplantation. In a particular embodiment, the method additionally comprises obtaining a biological sample from the patient = and analyzing the biological sample with an analysis program selected from the group consisting of Liquid chemical analysis, chromosomal translocation analysis, needle biopsy (needle b1〇: y), fluorescence in situ hybridization, laboratory biomarker analysis, immunostaining 4 dry staining, flow cytometry, or a combination thereof. For the purpose of naming 'the compound (4) Wei and ^ base components are numbered accordingly: 1 2 嘌呤

The "burning base" includes only the 163053.doc •35· 201242598 C and its linear, branched and cyclic monovalent hydrocarbon groups and combinations of such groups. Examples include mercapto, ethyl, isobutyl, cyclohexyl, cyclopentylethyl and the like. The total number of carbon atoms in each of these groups is sometimes described herein, e.g., when the group may contain up to 10 carbon atoms, it may be represented as 10C or C1-C10 or C1-10. "Halo" as used herein includes fluoro, ethane, bromine and iodine. Fluorine and gas are generally preferred. The term "selective ΡΙ3Κδ inhibitor" or "selective ΡΙ3Κβ inhibitor" as used herein refers to a compound which inhibits ρΙ3Κδ4ΡΙ3Κβ isozyme by inhibiting at least one other isozyme of the ΡΙ3Κ family, respectively. Selective inhibitors may also have activity against other isozymes of ΡΙ3Κ, but require a higher concentration to achieve the same degree of inhibition of other isozymes. "Selectivity" can also be used to describe a compound that is more inhibitory than a specific ΡΙ3 kinase than a similar agonist. "Selective ΡΙ3Κδ inhibitor" compounds are understood to be more selective for ΡΙ3Κδ than conventionally known as ΡΙ3Κ inhibitors. Compound (eg wortmannin or LY294〇02). At the same time, 渥曼青素 and LY294002 are considered as "non-selective PI3K inhibitors". Any type of compound that selectively negatively regulates the ΡΙ3Κδ expression or activity in certain embodiments can be used as a selective ΡΙ3Κδ inhibitor in the methods of the invention. In addition, any type of compound that selectively negatively regulates ΡΙ3Κδ expression or activity and has acceptable pharmacological properties can be used as a selective ΡΙ3Κδ inhibitor in the methods of treatment of the present invention. Without being bound by theory, the targeting of ρΐ 10δ inhibition by the compounds of the present invention provides a novel method for the treatment of hematological malignancies, in that the method inhibits constitutive signaling, thereby directly destroying the tumor 163053.doc -36 - 201242598 Tumor cells. Furthermore, without being bound by theory, inhibition of ρΐι〇δ will suppress microenvironmental signals that are critical for tumor cell homing, survival and proliferation. In an alternative embodiment, any type of compound that selectively negatively modulates (4) can be used as a selective ρΐ3κβ inhibitor in the methods of the invention. In addition, any type of compound that selectively negatively modulates beta expression or activity and has acceptable pharmacological properties can be used as a selective ρΙ3Κβ inhibitor in the methods of treatment of the present invention. "Treatment as used herein" means inhibiting a condition, i.e., arresting it; reducing the condition, even if it resolves; or ameliorating the condition, i.e., reducing the severity of at least one of the symptoms associated with the condition. In some embodiments, "treatment" refers to preventing a condition from occurring in an animal that has not been diagnosed with the condition. "Illness" is intended to cover, but is not limited to, medical conditions, diseases, conditions, syndromes and the like. In another aspect, the invention includes a method of inhibiting the function of a blood cell and/or mast cells and thereby being capable of treating a disease or condition characterized by hyperactivity or undesirable activity of a blood cell and/or mast cell. According to this method, the compound of the present invention which selectively inhibits the expression or activity of the fat-filling muscle SI3 kinase S (P(tetra)) in basophilic blood cells and/or mast cells can be used. Preferably, the method uses a P(10) inhibitor in an amount sufficient to inhibit the release of stimulated histamine by basophilic blood cells and/or mast cells. Thus, the use of such compounds and their fine KS selective inhibitors may have a therapeutic feature characterized by the release of histamine, i.e., allergic conditions including, for example, chronic obstructive pulmonary disease (C〇PD), asthma, ARDS, emphysema, and Diseases associated with the disease 163053.doc -37- 201242598 The value of the disease β method can be used to treat injuries that are susceptible or susceptible to reperfusion injury, that is, the period of time during which the tissue or organ undergoes ischemia and reperfusion. "Arts deduction of ischemia" refers to local tissue anemia caused by obstruction of arterial blood inflow. Transient ischemia and reperfusion will characteristically result in neutrophil activation and transmural transfer through the blood vessels into the affected area. The accumulation of activated neutrophils results in the production of reactive oxygen metabolites that damage the components of the tissues or organs involved. This "reperfusion injury" phenomenon is usually associated with conditions such as vascular stroke (including global cerebral ischemia and focal blood), hemorrhagic shock, myocardial ischemia or infarction, organ transplantation, and cerebral vasospasm. A illustrates that reperfusion injury occurs at the end of the cardiac bypass procedure or during a cardiac arrest when the heart begins to refill once the blood is blocked. It is expected that inhibition of Ρΐ3κδ activity will result in a decrease in the amount of reperfusion injury in these situations. In certain embodiments, the invention provides methods of treating a solid tumor. In a particular embodiment, the cancer is breast cancer, lung cancer, colon cancer or prostate cancer. In certain embodiments, the invention provides a method of treating a solid tumor associated with abnormal or undesirable cellular signaling activity mediated by ρΙ3Κβ: In some embodiments, the 'solid tumor line is selected from the group consisting of: Dirty cancer; bladder cancer, colorectal cancer; breast cancer, including metastatic breast cancer. Prostate cancer 'including androgen-dependent and androgen-independent prostate cancer, kidney cancer' includes, for example, metastatic renal cell carcinoma; hepatocellular carcinoma; lung cancer Including, for example, non-small cell lung cancer (NSCLC), bronchioloalveolar carcinoma (br〇nchi〇l (10) carcinoma 'BAC), and lung adenocarcinoma; ovarian cancer, including, for example, progressive 163053.doc -38- 201242598 skin cancer or primary Peritoneal cancer; cervical cancer; gastric cancer; esophageal cancer; head and neck cancer 'including, for example, head and neck squamous cell carcinoma; melanoma; neuroendocrine cancer' including metastatic neuroendocrine tumors; brain tumors including, for example, glial Gliooma, anaplastic oligodendroglioma, adult glioblastoma multiforme and Adult atypical stellate astrocytoma (aduh anaplastic astrocytoma); bone cancer; and soft tissue sarcoma. Genetic clearance ρΙΙΟδ has been found to produce a mild phenotype confined to the immune system. General observations include the organism's fertility without severe anatomy or behavioral abnormalities. Histological examination revealed that the main organs appeared to be normal. The total activity of class I ΡΙ3Κ in sputum cells and sputum cells was reduced by 30-50%. In addition, no increase in susceptibility was observed. In addition, the effects on the hematopoietic system include normal peripheral blood β β, lymphoid hypoplasia in the spleen and lymph nodes, and lack of germinal center and B220+IgM+ B cell progenitor cells in the bone marrow. The number is reduced, the serum immunoglobulin content is lowered, and the T cells in the thymus are developing normally. Genetic clearance of ρΙΙΟδ affects bone marrow and beta cell signaling, which play an important role in tumor formation. In particular, it affects the transmission, development, proliferation and survival of tyrosine kinase signals in bone marrow cells. The function of sputum cells is most affected and includes proliferation, differentiation, apoptosis and response to sputum cell survival factors (BCR, CD40, IL-4, chemokine). Accordingly, the present invention includes methods of treating abnormal or undesirable disease states of one or more of these bone marrow and B cell functions. At the molecular level, all of the ρΙΙΟα, ρΙΙΟβ, ρΙΙΟδ, and ρ11〇γ 163053.doc -39- 201242598 PI3K inhibitors (hvPS34, mTOR, DNA-ΡΚ and other inhibitors) were targeted to all tissues. Potential clinical indications include cancer, but clinically unfavorable events include hyperinsuljnemia in cancer patients. The advantage of dry-mediated inflammatory cells and cancer cells (where potential clinical indications include cancer, rheumatoid arthritis, asthma, allergies, and COPD) is that the treatment is well tolerated and avoids Such as the side effects of hyperinsulinemia. Thus, in one aspect, the invention provides a method of treating a patient suffering from insulin resistance or type 2 diabetes, cancer, rheumatoid arthritis, asthma, allergy, COPD or other conditions treatable with a compound of the invention method. For patients suffering from an over-insulin condition or propensity requiring such treatment, the compounds of the invention are particularly preferred over all-inhibitors. In certain embodiments, the compound of Formula I or Formula is preferred because it provides a therapeutic benefit in the treatment of hematological malignancies without adversely affecting insulin signaling. In one embodiment, the present invention relates to inhibition of ρΐ3Κ ρΙΙΟδ. The method. In another embodiment, the invention relates to a method of suppressing ρΐ3Κ ρΐ ι〇β or ρΙΙΟγ. In certain embodiments, the methods described herein have little or no off target aetivity. The method used in the specific method is for more than 3 protein kinases (including examples). The protein kinases summarized in Table 3 below showed little activity. In certain embodiments, the methods described herein do not have or have minimal hyperinsulinemia effects in cancer patients as compared to methods comprising administering a total ΡΙ3Κ inhibitor. In certain embodiments, the methods described herein are applicable to dry-mediated aging of 163053.doc 201242598 cells, since the compound of formula A inhibits Akt phosphorylation. Thus, a patient suitable for treatment with a compound of the invention may be selected, and in one embodiment, a patient exhibiting elevated Akt phosphorylation associated with a hematopoietic cancer, such as lymphoma, leukemia or multiple myeloma, is selected. The methods herein avoid the tendency to off target and are characterized by a negative result in the recipient gram screening, no hERG inhibition and no significant p45 inhibition. Another advantage of the method of the invention is that there are no adverse reactions in the cardiovascular, respiratory or central nervous system' as evidenced by the Institute of Safety Pharmacology. In addition, a 28-day toxicity study in rats and dogs proved to have a high therapeutic index, such as NOAEL (amount of no adverse reaction observed) > 1 〇μΜβ which is between the exposed group and its appropriate control group. The highest experimental dose of the chemical when there is no statistically significant or biologically significant increase in the frequency or severity of the rational effect. An adverse reaction is defined as any effect that results in a functional impairment and/or a pathological condition that affects the performance of the entire organism or reduces the ability of the organism to respond to another challenge. In certain embodiments, the methods described herein significantly alleviate lymph node lesions, thereby forcing lymphocytes into the circulation. This lymphocyte redistribution has the potential to place malignant CLL cells in a less protected environment outside the lymph nodes. In certain embodiments, co-administration of a chemotherapeutic and/or immunotherapeutic with a pharmaceutical agent of a compound of formula A enhances killing (:] [^ cells while reducing lymphocytosis. 'In another embodiment t' The method of the invention is not genotoxic in a standard set of tests. 163053.doc 41 201242598 Another advantage of the present invention is that the selectivity of the compound for one or two PI3K isoforms improves the safety profile over compounds with full PI3K inhibition. In another advantage, Compound I has a favorable target pharmacological profile with no good adverse effects on glucose or insulin content, and is well tolerated by normal healthy volunteers at doses above the usual therapeutic doses. Another advantage of the present invention includes the ability to treat a variety of hematological malignancies, as evidenced by the examples herein. In certain embodiments, the methods of the invention are directed to treating cancer. In certain embodiments, the cancer is a hematological malignancy. In the embodiment, the liquid cancer malignant tumor is selected from the group consisting of acute lymphocytic leukemia (ALL·), acute Myeloid leukemia (AML·), chronic lymphocytic leukemia (CLL·), multiple myeloma (MM), and non-Hodgkin's lymphoma (NHL·) 某些 In some embodiments, non-Hodgkin's lymphoma Choose from the following groups: diffuse large B-cell lymphoma (LDBCL), mantle cell lymphoma (MCL), Waldenstrom's macroglobulinemia (WM), and lymphoplasmacytic lymphoma. PI3K involves many Hematological malignancies and established pre-clinical proof of concept for treatment with Compound I. The following table summarizes specific hematological malignancies and methods for acting on primary patient cells or diseased cell lines. Primary role in patients with chronic lymphocytic leukemia (CLL) induces cell death and blocks survival factor 163053.doc -42- 201242598 Primary patient cells block PI3K signaling inhibition Proliferation cell line blocks PI3K signaling induction Apoptotic cell line blocks PI3K signaling and induces apoptosis. Primary cells in patients with 24/24 samples are overexpressed. 〇11〇δ induces apoptosis. Acute myeloid white blood (AML) acute lymphocytic leukemia (ALL)

Non-Hodgkin's Lymphoma (NHL) (MCL DLBCL, FL) Multiple Myeloma (MM) The information provided herein demonstrates that the compounds of the invention are useful in the treatment of lymphoma and leukemia. Lymphomas and leukemias usually selectively express P110 δ isoforms. For example, Figure 15 demonstrates that ρ 11 〇δ is dominant in most lymphoma cell lines, and ρ 11 〇α is usually not observed. In addition, the data presented in Figure 16 shows that cell cultures of six different leukemia cell lines are sensitive to Compound 1 and are strongly affected by this compound at a concentration of 5-10 micromolar. Figures 8 and 9 support Compound I in terms of reducing Akt (Ser473) production in several cell lines. For example, CLL mainly produces ριι〇δ and, to a lesser extent, Ρ11〇γ for signaling purposes, and thus it is expected that compounds inhibiting ρ110δ &/ or ρ11〇γ will exhibit selective cytotoxicity against such cells. Example 3 shows that Compound I has dose-dependent cytotoxicity against CLL cells (including cells from patients with poor prognosis (Figure 9) and cells showing resistance to other CLL therapeutics (Figure 20)) (Figure 3) ). In addition, Example 13 and Figure 13 demonstrate that administration of Compound 1 to a person at a rate of 50 mg BID during the 28 day period provides a significant therapeutic effect. A decrease in the percentage of ALC concentration in lymphocytes was observed. Thus, in one aspect, the invention provides a method of treating a CLL patient having drug resistant CLL using a compound of formula A 163053.doc -43 - 201242598. On the other hand, Example 丄7 shows that fibroblast cell lines that rely primarily on ρΙΙΟα for signal transduction are not sensitive to Compound I. Therefore, in one aspect, patient selection may include exclusion of cancers that are primarily dependent on ρΐ10α for signaling. The patient. The guanidine compounds are also suitable for the treatment of lymphomas, including both beta cell lymphoma and tau cell lymphoma. The data in Figure 4 demonstrates that six different ALL cell lines are sensitive to Compound I, which results in a significant decrease in cell viability of all six cell lines. Figure 12 and Example 12 demonstrate that on average, tumor burden was reduced by 44% in patients with mantle cell lymphoma treated with 50 mg of Compound I twice daily (BID) for 28 days. In addition, Figure 14 demonstrates that plasma compound I levels at the end of the 28-day period after administration of the 50 mg dose in MCL patients are similar to those observed in normal healthy volunteers (NHV); therefore, compounds are being treated There is no excessive accumulation during the cycle, and the patient does not become tolerant due to metabolic enhancement during the treatment cycle. Further, the compound of formula A or formula I is suitable for the treatment of hematopoietic cancers that constitutively exhibit Akt squamizing activity. Example 8 and Figures 8 and 9 list cancer cell lines showing constitutive Akt can acidification, including B cell lymphoma, tau cell lymphoma, ALL, malignant histiocytosis, DLBCL and AML » cell exposure Compound I reduces Akt phosphorylation. See also Example 19, which shows that Compound I inhibits constitutive Akt phosphorylation in 13/13 cell lines. In certain embodiments, the cancer is a solid tumor. In a particular embodiment, the cancer is breast cancer, ovarian cancer, lung cancer, colon cancer or prostate cancer. Figure 6 shows, for example, that Compound I reduced cell proliferation of two breast cancer cell lines, and Figure 163053.doc-44·201242598 10 illustrates cytotoxicity against three different breast cancer cell lines. Similarly, Figure 7 demonstrates that Compound I is cytotoxic to both ovarian cancer cell lines. For the treatment of solid tumors, it is advantageous to use a compound of formula A which exhibits a good activity against pU叩 (e.g., an IC50 of less than about i μΜ and preferably less than about 25〇nM_see Example 15), since solid tumors typically utilize this same The enzyme is not more than or more than 〇11〇δ. Thus, a compound of formula A having an IC50 of less than about 25 〇 nM is preferred for use in the treatment of solid tumors; the compound "", " is suitable for this use, as demonstrated herein. In some embodiments t, the individual to be treated is herein Described as an individual who has been diagnosed with at least one condition described herein that can be treated by the use of a compound of formula A. In some embodiments, the individual has been diagnosed with the cancers listed herein and has demonstrated at least one known Chemotherapeutic treatment is difficult to cure. For example, for treatment (such as proteasome inhibitors, autologous stem cell transplantation, CHOP regimen, rituximab, gasdabin, aletizumab, conventional anticancer Patients with non-reactive nucleoside analogs and alkylating agents) generally respond to the methods of treatment described herein. Thus, in one embodiment, the therapy of the invention is directed to a patient who has received one or more of the therapies In certain embodiments, the method of the invention is directed to a 3-cell or B-lymphocyte-associated disease. B cells play a role in the pathogenesis of autoimmune diseases. Formula A compounds (specifically I, I", Π and Π") are suitable for treating a plurality of individuals having a condition as described herein, particularly a human hematological cancer. In some embodiments, the individual selected for treatment of a hematological malignancy is an individual who relapses after other therapies 163053.doc -45 - 201242598 or is difficult to cure for other therapies. In some embodiments, the individual is selected to treat a hematological malignancy that is resistant to other cancer drugs. In some embodiments, the individual is selected to treat a hematological malignancy that exhibits a higher degree of activity. In some embodiments, the individual is selected to treat a hematological malignancy that exhibits a relatively low level of activity. In some embodiments, the individual is selected to treat a hematological malignancy that constitutively exhibits Akt phosphorylation activity. In one embodiment, the methods described herein comprise administering a compound of formula A as described herein in combination with a therapy for treating cancer. As used herein, "therapy" or "treatment" is the treatment of cancer by any of the well-known or experimental treatment forms that are used to treat cancer without the use of a compound of formula A. In certain embodiments, the combination of a compound of formula A with a conventional or experimental therapy for treating cancer or an autoimmune disease provides a benefit and/or desired therapeutic result that is superior to the result of not using the combination therapy. . In certain embodiments, therapies for treating cancer are well known to those of ordinary skill and are described in the literature. Therapeutic treatments include, but are not limited to, chemotherapy, combinations of chemotherapy, biological therapy, immunotherapy, radioimmunotherapy, and Use monoclonal antibodies and vaccines. In some of the above embodiments, the combination method provides for the simultaneous administration of a compound of the formula and administration during the administration period of the therapy. In some of the above embodiments, the guanidine compound is administered concurrently with other chemotherapeutic therapies. In certain embodiments, the combination method dictates that the compound of formula A is administered prior to or after administration of the therapy. In some of the above embodiments, the system is at least one standard or experimental 163053.doc -46-201242598 chemotherapy is difficult to cure. In some of the above embodiments, the system is difficult to cure with at least two standard or experimental chemotherapies. In some of the above embodiments, the system is difficult to cure with at least three standard or experimental chemotherapies. In some of the above embodiments, the system is difficult to cure with at least four standard or experimental chemotherapy therapies. In some of the above embodiments, the M solid is at least one standard or experimental chemotherapeutic selected from the group consisting of: fludarabine, rituximab, sieving agent, alenizumab And the above-mentioned chemotherapeutic aq 〇 In some of the above embodiments, the 'system is at least two standard or experimental chemotherapeutic treatments selected from the group consisting of: m. fludarabine, rituximab, and thiol The agent, alenizumab, and the chemotherapeutic remedies listed above are in some of the above embodiments, the system being at least three standard or real purification therapies selected from the group consisting of: (4): gas Dalazine, rivastigmine, avidin, alenizumab, and the chemotherapeutics listed above in the above-mentioned embodiments, the system is at least four selected from the group consisting of Or experimental chemotherapy is difficult to treat n Darabine, Lee

Tetuximab, calcinating agent, alum, nD J丨7, early resistance to the above-listed chemotherapy aq 确切 The exact details of the individual morbidity of the combination of the drug and the selected therapy can be determined experimentally. . The administration of formula A is compounded; the improvement of the order and timing of the employee should be appropriate to the judgment of the individual, the subject, and generally the attending physician. 163053.doc -47· 201242598 Non-limiting examples of experimental or standard therapies are described below. In addition, the treatment of certain lymphomas is reviewed in the following: Cheson, BD, Leonard, JP, "Monoclonal Antibody Therapy for B-Cell Non-Hodgkin's Lymphoma" The New England Journal of Me Mountain 2008, 359(6), Pp. 613-626; and Wierda, WG, "Current and Investigational Therapies for Patients with CLL" Hematology 2006, pp. 285-294. Morton, L. M., et al., "Lymphoma Incidence Patterns by WHO Subtype in the United States, 1992-2001" 2006, 107(1), pp. 265-276, analyzes the profile of the onset pattern of lymphoma in the United States. Treatment of non-Hodgkin's lymphoma, especially non-Hodgkin's lymphoma of B cell origin including, but not limited to, the use of monoclonal antibodies, standard chemotherapy (eg CHOP, CVP, FCM, MCP and the like), Radioimmunotherapy and combinations thereof, especially the integration of antibody therapy with chemotherapy. Non-limiting examples of unconjugated monoclonal antibodies for non-Hodgkin's lymphoma/B cell cancer include rituximab, aleuzumab, human or humanized anti-CD20 antibodies, luciximab, Anti-TRAIL, bevacizumab, galiximab, irradizumab, SGN-40 and anti-CD74. Non-limiting examples of experimental antibody agents for treating non-Hodgkin's lymphoma/B cell cancer include olfaximab, ha20, PR0131921, aleuzumab, glipizumab, SGN-40 , CHIR-12.12, irradiizumab, luciximab, apozumab, milatuzumab and bevacizumab. Any monoclonal antibody can be combined with rituximab, fludarabine or a chemotherapeutic/therapy. 163053.doc • 48 - 201242598 Non-limiting examples of standard chemotherapy for non-Hodgkin's lymphoma/B cell cancer include CHOP (cyclophosphamide, cranberry, vincristine, prednisone), FCM (fludarabine, cyclophosphamide, mitoxantrone), CVP (cyclophosphamide, vincristine and prednisone), MCP (mitoxantrone, phenylbutyrate and chlorpyrifos) R-CHOP (rituximab + CHOP), R-FCM (rituximab + FCM), R-CVP (rituximab + CVP) and R-MCP (R-MCP). Non-limiting examples of radioimmunotherapy for non-Hodgkin's lymphoma/B cell cancer include 钇-90 labeled temtanisobezumab and iodine-131 labeled tocilizumab. These therapeutic agents are approved for use in individuals with relapsed or refractory vesicular lymphoma or low grade lymphoma. Therapeutic therapies for mantle cell lymphoma include combination chemotherapy, such as CHOP (cyclophosphamide, cranberry, vincristine, prednisone), multi-fraction CVAD (hyperfractionated ring decylamine, Changchun new test, cranberry, dexamethasone, amidoxime, cytarabine) and FCM (fludarabine, cyclophosphamide, mitoxantrone). In addition, these therapies may be supplemented with the monoclonal antibody rituximab (Rituxan) to form combination therapy R-CHOP, multi-fraction CVAD-R and R-FCM. Other methods include combining any of the above therapies with stem cell transplantation or ICE (iphosphamide, carboplatin and etoposide) therapies. Another method of treating mantle cell lymphoma includes immunotherapy, such as the use of monoclonal antibodies, such as rituximab (Rituximab). Rituximab is also effective against other inert B cell cancers, including marginal lymphoma, WM, CLL, and small lymphocytic lymphoma. The combination of rituximab and a chemotherapeutic agent is particularly effective. 163053.doc -49- 201242598 It is effective. One improved method is radioimmunotherapy, in which monoclonal antibodies are combined with radioisotope particles such as iodine-131 tosimolumab (Bexxar®) and technetium-90 temtanisobezumab (Zevalin®). In another example, Bexxar® and CHOP are used sequentially in therapy. Another example of immunotherapy involves the use of a cancer vaccine based on the genetic makeup of individual patient tumors. An example of a lymphoma vaccine is GTOP-99 (MyVax®). Another method of treating mantle cell lymphoma involves autologous stem cell transplantation combined with high dose chemotherapy. Another method of treating mantle cell lymphoma involves administration of a proteasome inhibitor such as Velcade® (bortezomib or PS-341); or an anti-angiogenic agent such as thalidomide, especially in combination with rituximab. Another treatment is the administration of a drug that causes degradation of the Bcl-2 protein and increases the sensitivity of the cancer cell to chemotherapy, such as Olimpson (Genasense) in combination with other chemotherapeutic agents. Another method of treatment involves administering an mTOR inhibitor that causes inhibition of cell growth and even cell death; a non-limiting example is sirolimus (CCI-779), and sirolimus and Rituxan®, Velcade ® or a combination of other chemotherapeutic agents. Other recent therapies for MCL have been revealed (iWziwre / Reviews; Jares, P. 20〇7). Non-limiting examples include Flavopiridol, PD0332991, R-roscovitine (Selicilib, CYC202), Styrene oxime, Obatoclax (GX15-070) ), TRAIL, anti-TRAIL DR4 and DR5 antibodies, sirolimus (CC1-779), everolimus (RAD001), BMS-345541, curcumin, volcanum (SAHA), Shali Degree amine, lenalidomide 163053.doc -50- 201242598 (vlimid, CC-5013) and geldanamycin (17-AAG) 〇 for the treatment of Waldenstrom's macroglobulinemia Non-limiting examples of other therapeutic agents include parisone, bortezomib (Velcade®), rituximab, sildenafil citrate (Viagra®), CC-5103, thalidomide Ira Tocilizumab (hLL2-antiC: D22 humanized antibody), simvastatin/T heart flolin, capsi-iHOampath-lH), dexamethasone, DT PACE Olimpson, anti-Nipula A10, anti-Xinpratong AS2-1, aleizumab, beta alixine, cyclophosphamide, cranberry hydrochloride, prednisone, vincristine sulfate, fludarabine, Gestrin, melphalan, recombinant interferon alpha, carmustine, cisplatin, cyclophosphamide, cytarabine, etoposide, melamine, dolastatin 1 〇, indium Ι 丨Antibody ΜΝ _ 14 , 钇Υ 90 humanized erarazumab, anti-thymocyte globulin, busulfan 'cyclosporine, f-amine oxime, mycophenolate morpholine ethyl ester, therapeutic allogeneic lymphocytes , 钇γ 90 temtanisobezumab, sirolimus, tacrolimus, carboplatin, thiotepa, paclitaxel, adiponectin, recombinant interferon alpha, taxol, isoxanthin Amine, Mesna, Recombinant Interleukin-12, Recombinant Interleukin_u, Bci_2 Family Protein Inhibitor αβτ_ 263, Tunicin Fusion Toxin, Tanspirin, Everolimus, PEGylation Filgrastim, vorinostat, avostatin, recombinant flt3 ligand, recombinant human thrombopoietin, lymphokine-activated killer cells, amine-filled/butyrate 5, amine camptothecin, irinote Conhydrochloride, caspofungin acetate, clolapabine, afarbein, naribine, pentastatin, sax , vinorelbine tartrate hydrogen salt, | Ding-1 analog peptide vaccine, Jie 163053.doc -51 · 201242598 126-134 peptide vaccine, retinoid, Ixabepilone, (four) force # strain antibody CD19, single branch Early strain antibody CD20, omega-3 fatty acid, mitoxantrone salt, oleic acid acetate, tosimizumab and moth 1-131 tosimozumab, momi, dioxon, typography Farnese, autologous human tumor-derived HSPPC_96, veltuzumab, bryostatin 1 and pegylated liposomal cranberry hydrochloride and any combination thereof. Non-limiting examples of other therapeutic agents for the treatment of diffuse large b-cell lymphoma (DLBCL) (Others i/ 2〇〇5 Abrams〇n,j) include cyclophosphamide, cranberries , vincristine, prednisone, anti-CD2 〇 monoclonal antibody, etoposide, bleomycin, many of the agents listed for Waldenstrom's macroglobulinemia and any combination thereof, such as ICE and RICE. Non-limiting examples of therapeutic procedures for treating Waldenstrom's macroglobulinemia include peripheral blood stem cell transplantation, autologous hematopoietic stem cell transplantation, autologous bone marrow transplantation, antibody therapy, biological therapy, enzyme inhibitors Therapy 'body irradiation, stem cell infusion, bone marrow removal under stem cell support, peripheral blood stem cell transplantation in vitro, cord blood transplantation, immunoenzymatic technology, pharmacological studies, low LET cobalt-60 gamma ray therapy, bleomycin, ha Know surgery, radiation therapy and non-myeloablative allogeneic hematopoietic stem cell transplantation. Non-limiting examples of other therapeutic agents for the treatment of chronic lymphocytic leukemia (Spectrum, 2006, Fernandes, D_) include phenylbutyrate (Leukeran), cyclophosphamide (Selone ( Cyl〇xan), Endoxan, Endoxana, Cyclostin, Fludarabine (Fludara), Penstatin (Nipent) ), 163053.doc -52· 201242598 Clarendam (Leustarin), Cranberry (AdHamycin®, Adriblastine), Vincristine (Vincristine) ), Penny's privately-loved nylon, aleuzumab (MabCampath), many agents and combination chemotherapy for Waldenstrom's macroglobulinemia and Chemical immunotherapy, including common combination therapies: cvp (cyclophosphamide, vincristine, prednisone); R_cvp (rituximab-CVP); ICE (isoprene, carboplatin, etoposide ); R_ ICE (rituximab·ICE); FCR (fludarabine, cyclophosphamide, rituximab); and FR (fludarabine, rituximab) Monoclonal antibody). In certain embodiments, the methods comprise administering to the patient, in addition to the compound, a therapeutically effective amount of at least one therapeutic agent and/or therapeutic procedure selected to treat the cancer of the patient. In certain embodiments, the method comprises administering to the patient, in addition to Compound I or II, a therapeutically effective amount of a combination of therapeutic agents selected from the group consisting of: a) bendamustine; b) Infliximab; c) bendamustine and rituximab; sentence olfaximab; and seletonide. The compounds of the present invention can be formulated for administration to individual animals using formulation techniques well known in the art as generally known. Formulations suitable for specific modes of administration and formulas of ruthenium compounds can be found in Remingt〇n, s pharmaceutica|

Sciences, latest edition, Mack Publishing Company, Easton, PA. The compounds of the invention may be prepared in the form of a prodrug, i.e., the protected form of the compound of the invention is released upon administration to an individual. Typically, the protecting group is hydrolyzed in a body fluid, such as in an iL stream, to release the active compound, or a protecting group. 163053.doc • 53· 201242598 is oxidized or reduced in vivo to release the active compound. A discussion of prodrugs can be found in Smith and Wmiams Introduction to the Principles of Drug Design, Smith, H.J.; Wright, 2nd edition, London (1988). The compounds of the invention may be administered in purely chemical form, but it is generally preferred to administer the compound in the form of a pharmaceutical composition or formulation. Accordingly, the present invention also provides a pharmaceutical composition comprising a compound of formula A and a biocompatible pharmaceutical carrier, adjuvant or vehicle. The composition may include, as the sole active moiety, a combination of a compound of formula A with other agents (such as oligonucleotides or polynucleotides, oligopeptides or polypeptides, drugs or hormones), with excipients or other pharmaceutically acceptable The carrier mixture can be considered pharmaceutically acceptable insofar as the carrier and other ingredients are compatible with the other ingredients of the formulation and are not deleterious to the recipient. The pharmaceutical compositions may be formulated to contain a pharmaceutically acceptable carrier and, if appropriate, excipients and auxiliaries which facilitate the processing of the active compound into preparations which can be used in the pharmaceutical compositions. The form of administration will usually determine the nature of the carrier. For example, a formulation for parenteral administration may comprise an aqueous solution of the active compound in a water-soluble form. A carrier suitable for parenteral administration may be selected from the group consisting of physiological saline, buffered saline, dextrose, water, and Other Physiologically Compatible Solutions "The preferred carrier for parenteral administration is a physiologically compatible buffer, such as Hank's solution, Ringer's solution or physiology. Buffer saline. For tissue or cell administration, penetrants appropriate to infiltrate specific barriers are used in the formulation. Such penetrants are generally known in the art. For formulations containing proteins, the formulations may include stabilizing materials such as polyols (e.g., sucrose) and / 163053.doc • 54-201242598 or surfactants (e.g., nonionic surfactants) and the like. Alternatively, the formulation for parenteral administration may comprise a dispersion of the active compound or a suspension prepared as a suitable oily suspension. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil and synthetic fatty acid esters such as ethyl oleate or triglycerides or liposomes. The aqueous injectable suspension may contain a substance which increases the viscosity of the suspension such as carboxy-methylcellulose sodium, sorbitol or dextran. The suspension may optionally contain an agent suitable for stabilizing or increasing the solubility of the compound to allow for the preparation of a high concentration solution. Aqueous polymers which provide pH sensitive dissolution and/or sustained release of the active agent can also be used as a coating or matrix structure, such as a mercaptopropionate polymer, such as

Eudragit® series from Rohm America Inc. (Piscataway, N.J.). Emulsions such as oil-in-water and water-in-oil dispersions may also be used, optionally emulsified or dispersing agents (surfactants; surfactants). The suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, Gum tragacanth and mixtures thereof. Liposomes containing the active compound of formula A can also be used for parenteral administration. Liposomes are usually derived from phospholipids or other lipid materials. Compositions in liposome form may also contain other ingredients such as stabilizers, preservatives, excipients, and the like. Preferred lipids include natural and synthetic phospholipids and phospholipids (lecithin). Methods of forming liposomes are known in the art. See, for example, Prescott (ed.), Methods in Cell Biology, Volume XIV, page 33, Academic Press, New York (1976). 163053.doc • 55- 201242598 A pharmaceutical composition comprising a compound of formula A suitable for oral administration can be formulated using a pharmaceutically acceptable carrier well known in the art. The preparation for oral administration may be in the form of a key, a pill, a capsule, a cachet, a dragee, a lozenge, a liquid, a gel, a syrup, a serum, an elixir, a suspension or a powder. For the purpose of illustration, a pharmaceutical preparation for oral use can be obtained by combining the active compound with a solid excipient, optionally grinding the resulting mixture and, if necessary, processing the mixture of granules after adding suitable auxiliaries to obtain a sharp or sugar-coated pill. core. Oral formulations may employ a liquid carrier of a type similar to that used parenterally, such as aqueous buffer solutions, suspensions, and the like. Preferred oral formulations include lozenges, dragees, and gelatin capsules. Such formulations may contain one or more excipients including, without limitation: a) a diluent such as a sugar, including lactose, dextrose, sucrose < mannitol or sorbitol; b) a binder, such as Magnesium aluminum silicate, starch from corn, wheat, rice potato, etc.; e) Cellulosic materials such as methylcellulose, (tetra)methylcellulose, and sodium sulphonate, polyethylene. More than (iv) ketones, gums (such as acacia and tragacanth) and proteins (such as gelatin and collagen); d) disintegrants or solubilizers such as cross-polypyrrolidinium, starch, agar, (tetra) acid Or a salt thereof (such as sodium) or a pure composition; a moisturizing agent such as sulphur dioxide, talc, stearic acid or its salt or calcium salt, and polyethylene glycol; f) seasoning And sweeteners; 163053.doc -56- 201242598 g) colorants or pigments, for example to identify products or to characterize the amount of active compound (dosage); and h) other ingredients such as preservatives, stabilizers, bulking agents, An emulsifier, a dissolution promoter, a salt for regulating osmotic pressure, and a buffer. In some preferred oral formulations, the pharmaceutical composition comprises at least one selected from the group (a) above, or at least one selected from the group (b) above, or at least one selected from the group consisting of a substance, or at least one selected from the group (d) above, or at least one selected from the group (e) above. Preferably, the composition comprises at least one substance selected from each of the two groups of groups (a) - (e) above. Gelatin capsules include push-fit capsules made of gelatin and soft seal capsules made of gelatin and a coating such as glycerol or sorbitol. The capsule may contain the active ingredient in admixture with a filler, a binder, a lubricant, and/or a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable fluid, such as a fatty oil, liquid paraffin or liquid polyethylene glycol, with or without a stabilizer. The dragee pill may have a suitable coating, such as a concentrated sugar solution, and the coating may also contain gum arabic 'talc, polyethylene ratio, brewing J, erbopol gel, polyethylene glycol. And / or titanium dioxide, coating solutions and suitable organic solvents or solvent mixtures. The pharmaceutical composition can be provided as a salt of the active compound. The salt is more soluble in aqueous or other protic solvents than the corresponding free acid or free base form. Pharmaceutically acceptable salts are well known in the art. The compound containing an acid moiety can form a pharmaceutically acceptable salt of 163053.doc -57- 201242598 with a suitable cation. Suitable pharmaceutically acceptable cations include, for example, alkali metal (e.g., sodium or potassium) and alkaline earth (e.g., calcium or magnesium) cations. A compound of the formula (Α) containing an illustrative moiety can form a pharmaceutically acceptable acid addition salt with a suitable acid. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J/^(10) 66:1 (1977). Salts can be prepared in situ during the final isolation and purification of the compounds of the invention or by reacting the free base functional groups with a suitable acid. Representative acid addition salts include, but are not limited to, acetates, adipates, alginates, citrates, aspartates, benzoates, phthalates, hydrogen sulphates, Butyrate, camphorate, camphorolsulfonate, digluc〇nate, glycerol phosphate, hemisulfate, heptanoate, hexanoate, fumarate, Hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethane sulfonate (isosulfonate), lactate, maleate, methanesulfonate or sulfate, niacin Salt, 2-naphthalene sulfonate, oxalate, pamoate, pectate, peroxosulfate, 3-propionic propionate, picrate, pivalate, C Acid salts, succinates, tartrates, thiocyanates, phosphates or hydrogen citrates 'glutamate, hydrogencarbonate, p-toluenesulfonate and undecanoate. Examples of acids which can be used to form pharmaceutically acceptable acid addition salts include, without limitation, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and acid loading; and organic acids such as oxalic acid, maleic acid, and butyl. Diacid and citric acid. The inert gas-containing groups can be quaternized with a reagent such as a lower alkyl halide such as methyl, ethyl, propyl and butyl chloride, bromide and telluride; Dimethyl sulphate, diethyl ester, dibutyl ester 163053.doc • 58 · 201242598 and monoamyl ester; long chain alkyl halides, such as odor, __, turf, turf, and fifteen An alkylate, a desertification, and a Wei; an aryl radical, such as benzyl and phenethyl desert; or other agents. Thereby, a product which is improved in solubility or dispersibility is obtained. A plant comprising a compound of the invention formulated in a pharmaceutically acceptable carrier can be prepared, placed in an appropriate container t, and labeled for treatment of a condition indicative. And ' also encompasses an article, such as a dosage form comprising a compound of the invention and a container containing the instructions for use of the compound. The invention also encompasses a cover kit. For example, a kit can comprise a dosage form of a pharmaceutical composition and a package insert containing instructions for using the composition to treat a medical condition. In either case, the condition indicated on the label can include treatment of an inflammatory condition, cancer, and the like. Methods of Administration Pharmaceutical compositions comprising a compound of formula A can be administered to an individual by any conventional method, including parenteral and enteral techniques. Parenteral administration forms include compositions by administration other than via the gastrointestinal tract, such as intravenous, intravascular, intraperitoneal, intramedullary, intramuscular, intra-articular, intrathecal, and intraventricular injections. Enteral administration forms include, for example, oral (including buccal and sublingual) and rectal administration. Transdermal administration forms include, for example, transmucosal administration and transdermal administration. Transmucosal administration includes, for example, enteral administration as well as nasal, inhalation, and deep lung administration; vaginal administration; and rectal administration. Transdermal administration includes passive or active transdermal or transcutaneous forms including, for example, patch and iontophoresis devices, as well as topical application pastes, ointments or ointments. Parenteral administration can also be achieved using high pressure techniques such as 163053.doc •59· 201242598 powderjecttm. Surgical techniques include implantation of a reservoir (like, a preferred administration of w:: = local or surface delivery or delivery to a dispersive condition, such as for reperfusion injury or for systemic conditions (such as 2 (sept) 〗 〖Cemla)) intravenous transmission. For diseases of the respiratory tract, such as chronic obstructive pulmonary disease,:: disease, including 丞f lung disease, asthma and emphysema, administration of inhalation or occupation and spray _, aerosol In addition to the above-mentioned actual wipes, the compound of formula A is administered during or after administration of chemotherapy, radiation therapy and/or surgery. Selected formulations and routes of administration It should be appropriate for the individual individual, the condition of the individual being treated, and generally the judgment of the attending physician. The therapeutic index of the compound of formula A can be enhanced by modifying or derivatizing the compound for targeted delivery to cancer cells that express the marker. Identifying a cell as a cancer cell. For example, the compound can be linked to an antibody that recognizes a marker that is selective or specific for cancer cells so that the compound reaches the vicinity of the cell to locally exert it. Used as previously described (see, for example, Pietersz, et al., Wimoi Λβν, 129:57 (1992); Trail, et al., > Scze«ce, 261:212 (1993); and R〇wlinson-Busza, Et al., Cwrr Ortco/, 4:1142 (1992). The targeted delivery of tumors of these compounds enhances the therapeutic benefit', especially minimizing potential non-specific toxicity due to radiation therapy or chemotherapy. In the sample, the compound of formula A and the radioisotope or chemotherapeutic agent can be combined with the same anti-tumor antibody. 163053.doc • 60 - 201242598 The characteristics of the agent itself and the formulation of the drug can be used to control the physical state and stability of the agent; t, in vivo release rate, and in vivo clearance rate. This pharmacokinetic and pharmacodynamic information can be collected via preclinical in vitro and in vivo studies 'subsequent confirmation in humans during clinical trials. ^ Thus, for the method of the invention The therapeutically effective dose of any compound used in the towel can be estimated initially by biochemical and/or cell-based analysis. Next, the animal model can be used to dose to achieve specific pl3K isoforms. The desired circulating concentration range for the performance or activity of a combination of isoforms or isoforms. As human studies progress, further information regarding the appropriate dosage and duration of treatment for various diseases and conditions will be obtained. Good sexuality, but one of the limitations on the therapeutic dose (4) elevated liver function test (LFT). LFT involves standard clinical biochemical tests on the patient's serum or plasma to provide information about the patient's liver status. Exceeding the normal range, such as alanine transaminase, aspartate transaminase, alkaline phosphatase, bilirubin & glutamate thiol transpeptidase, may indicate possible hepatotoxicity. The dose of the therapeutic compound can be adjusted to avoid or reduce the likelihood of elevated liver function test values and subsequent hepatotoxicity. For example, an incremental dose of a compound can be administered to an individual. At a certain dose, the individual begins to show an increase in the LFT level beyond the normal range, indicating that hepatotoxicity may be present under the agent. In response, the dosage can be reduced to a level that reduces the LFT level to an acceptable level as judged by the treating physician, e.g., within a range normal to the individual being treated or within about 25% to 50% of the normal level. The standard. Therefore, liver function tests can be used to titrate the dose of a compound. 163053.doc -61 - 201242598 The toxicity and therapeutic efficacy of these compounds can be determined in cell cultures or experimental animals by standard pharmaceutical procedures, such as determining LD5〇 (a group of lethal doses) and ed50 (in groups) 50% of the doses with therapeutic effectiveness). The dose ratio between the toxic effect and the therapeutic effect is "therapeutic index, which is usually expressed as the ratio LD50/ED50. It exhibits a larger therapeutic index, that is, a compound having a sputum dose substantially higher than the effective dose. From this cell culture Information obtained from analysis and other animal studies may be used to establish dose ranges for human use. The doses of these compounds are preferably within the circulating concentration range of edm including little or no toxicity. The dose may be limited by treatment-related toxicity. Symptoms. In addition to elevated liver function tests, these symptoms also include anemia 'sight blur, diarrhea, vomiting, fatigue, mucositis, peripheral edema, fever, peripheral neuropathy, pleural effusion, night sweat (night(10)(10) And orthopnea or a combination thereof, at a dose, if the individual develops such symptoms of intolerance, the dose can be lowered to eliminate adverse events and no longer have adverse events, or reduced to treatment Acceptance as judged by the physician" Another consideration in determining the dose of a compound for 6 patients is in the plasma. The desired concentration. In a particular embodiment, the concentration of the compound in the blood is between 4〇3 〇〇〇ng/mL2 during the hour from the administration. In another specific embodiment, from the time of administration During the 12-hour period, the concentration of the compound in the blood is between 75-2,000 ng/mL. In another specific embodiment, 'from the time of self-administration! 2 hours _, the concentration of the compound in the blood is between 500-2,000 ng Between /mL. In a preferred embodiment, during the 12 hours from the 163053.doc •62·201242598 drug, the concentration of human in the ash is between 40-3,000 ng/mL. Wherein the compound is 1, 11 or II and the oral dose is about, (10) mg '15 〇 mg or _. In the preferred embodiment, 12 small _ from the time of administration, the compound indifference in the liquid is between 40-3, between hydrazines, wherein the compound is of formula I and the oral amount is about 50 mg, _mg, 150 mg or mg. In a preferred embodiment, the 12-hour period fa1 'blood from the time of administration The concentration of the compound is between 40-3,000 ng/mL, where the compound is of the formula „and the oral dose is about 5〇, ι〇〇mg, 150 mg or 200 mg.In some of the above embodiments, the maximum concentration in plasma is achieved within two hours of administration. In certain embodiments, the dosage of the compound or formula is selected to produce an average drug plasma concentration of about 10 11 or more or more than 1 〇 nM over a period of 8 to 12 hours, and provides about 500 nM or more. Preferably, the peak plasma concentration is above about 1000 nM or 1000 η. In certain embodiments, the dose of the compound of formula II is selected to produce an average drug of about 1 〇〇 11] or more than 100 nM over a period of 8 to 12 hours. Plasma concentration, and provides a peak concentration of about 5 〇〇 nM or more, preferably about 1 〇〇〇 nM or more than 1000 nM. In certain embodiments, 'select a compound of formula I or formula II The dose produces an average drug blood test concentration of about 200 nM or more over a period of 8 to 12 hours and provides a peak plasma concentration of about 500 nM or more, preferably about 1 〇〇〇 nM or more. In certain embodiments, a dose of a compound of Formula I or Formula II is selected to produce a plasma concentration within a range of observed compound effects that are observed for therapeutic effects, such as apoptosis of cancer cells. In certain embodiments, selection Formula I or Formula II 163053.doc •63- 201242 The dose of the 598 compound is such as to produce a plasma concentration equal to or higher than the pH of the ρΙ3κδ isoform activated ECso in the plasma. In certain embodiments, the dose of the compound of formula or formula II is selected to be at least from the self-administered compound During the 12-hour period, the ECw content was higher than the ΡΙ3Κδ activation ECw content in the cells and lower than the ΡΙ3Κγ activation £ (:5〇 content. For example, if the ECso value of 嗜^^^ basophilic blood cell activation in whole blood plasma The 65w η Μ and the ECw value for activation of ΡΙ 3 Κ basophilic hematocrit is 1100 η Μ, and the selected compound dose yields the lowest compound plasma concentration between 60 η Μ and 11 〇〇 Μ 8 from 8-12 hours from the administration of the compound. Similarly, the dose can be selected such that the minimum blood concentration is higher than the ecm content that activates ρπκδ basophilic blood cells and is lower than the ECw content that activates ρΐ3Κα, ρΐ3Κβ or ΡΙ3Κγ basophilic blood cells. In vivo activation of ρΐ3Κ isoforms Or the ECw value of inhibition can be determined by a general practitioner. In an alternative embodiment, the upper limit of the minimum drug concentration may exceed ΡΙ3Κγ, ΡΙ3Κα, or ΡΙ3Κβ in plasma. The EC5 depreciation of isoforms is not limited. In addition, the blood concentration range of the drug is at a therapeutic level in the treatment of hematological malignancies while minimizing adverse side effects. For example, 'although with δ Selective, but the compound exhibits sufficient activity for p 1 丨〇 γ to be clinically useful, that is, effective for cancers that rely on ρ11 〇 γ to achieve k-transduction', because it is possible to achieve a higher than effective inhibition. The plasma content of the dose is 'selective relative to other isoforms, specifically alpha isoforms. Thus, in some embodiments, the compound dose is selected to produce an effective selective inhibition of the blood concentrations of ρ11〇5 and ρ11〇γ. 163053.doc • 64· 201242598 In some embodiments the dose of the compound resulted in a minimum plasma concentration between 65 nM and 11 〇〇 nM during the 8 to 12 hours from the administration of the compound. In some of the above embodiments, the period of time is at least 12 hours from the administration of the compound. In a particular embodiment, the compound is administered in a therapeutically effective amount. In a particular embodiment, the compound is administered at a dose of from 20 to 500 mg/day. In a particular embodiment, the compound is administered at a dose of 50-250 mg/day. 8 In a particular embodiment, the compound is administered at a dose of 25 mg to 150 mg per dose, and two doses are administered per day ( For example, it is administered twice a day at a dose of 25^^ to 〇5〇mg). In a preferred embodiment, the individual is treated twice daily with 5 〇 ^^ to 丨〇〇 mg of the compound of formula A. In other preferred embodiments, the individual is treated twice daily with 150 mg of the compound of formula A. In a particular embodiment, the method comprises administering to the patient an initial dose of from 20 to 500 mg of the compound per dose and incrementing the dose until clinical efficacy is achieved. The dose escalation can be in increments of about 25, 50, 100 or 150 mg. Dosage can be increased every week, every other day, twice a week or once a week. In a particular embodiment, the method comprises continuing to treat the patient by administering the same dose of the compound to achieve clinical efficacy or by incrementally decreasing the dose to a level at which efficacy can be maintained. In a specific embodiment, the method comprises administering to the patient an initial dose of 20-500 mg per dose and increasing the dose to a total daily dose of 50-400 mg during at least 6 days » the dose may be further increased to about 75 〇mg/day 163053.doc -65- 201242598 In a particular embodiment, the compound is administered at least twice daily. In a particular embodiment, the compound is administered orally, intravenously or by inhalation. Preferably, the compound is administered orally. In some embodiments, oral administration of beta at a dose of about 50 mg BID, about 100 mg BID, or about 150 mg BID. For the methods of the invention, any effective dosing regimen of prescribed dosage schedules and sequences can be used. Preferably, the dosage of the medicament comprises a unit of medical dosage comprising an effective amount of the medicament. As used herein, "effective amount" refers to an amount sufficient to modulate ΡΙ3Κδ expression or activity and/or obtain a measurable change in an individual's physiological parameter via administration of one or more medical dosage units. "Effective amount" can also mean the amount needed to improve an individual's disease or condition. The suitable dosage range for the guanidine compound varies depending on such considerations, but in general, the compound is administered in a range of 1 〇.〇 μβ·15 mg; 1.0 mg or 0.5 mg-5 mg per kg body weight. Thus, for a typical 7 〇 kg human subject, the dose range is 700 pg to 1050 mg per dose; 70 pg JOO mg; or 35 mg to 3 50 mg, and two or more doses may be administered per day. The dose may be higher when the compound is administered orally or transdermally, e.g., intravenously. A reduced toxicity of the guanidine compound allows for a therapeutically higher dose. In some of the above embodiments, up to 75 mg/day of the compound of the invention is orally administered. In some of the above examples, the compound of formula A was administered at a dose of 50 mg BID. In some of the above examples, the Formula A compound was administered at a dose of 100 mg BID. In some of the above examples, the compound of formula A is administered at a dose of 150 mg BID. In some of the above examples, the compound of formula A was administered at a dose of 200 mg BID. In the above example 163053.doc • 66· 201242598, the compound of formula A was administered at a dose of 350 mg BID. In a particular embodiment, for the treatment of leukemia, lymphoma and multiple myeloma, a dose of about 50-350 mg per dose per day is usually administered orally or preferably twice a day. In some of the above embodiments, oral administration of up to 750 mg/day of Compound I or II" is suitable. In some of the above embodiments, the formula " or formula H" compound is administered at a dose of 5 〇 mg BID. In some of the above embodiments, the compound of formula, or formula II" is administered at a dose of 1 〇〇 mg BID. In some of the above examples, the formula I" or the hydrazine compound is administered at a dose of 150 mg BID. In some of the above examples, the compound of formula I or formula II' is administered at a dose of 2 〇〇 mg bid. In some of the above examples, the formula is " or formula π, the compound is administered at a dose of 35 〇. Mg BID administration. In some of the above examples, for the treatment of leukemia, lymphoma, and multiple myeloma, each dose of about 50.35 〇m η" compound is administered orally once or preferably twice a day is often suitable: The compound can be administered in a single bolus dose, such as intravenous or transdermal administration over a period of time, or in multiple doses. Administration continues for at least one cycle. In some embodiments, the cycle is at least 7 days. In some embodiments, the period is about (4). In some embodiments, for (4) 28 days and then discontinued for at least n of the embodiments, the complete period is 28 days of continuous daily administration. After each period Measurable evaluation of clinical response to patients 1 bed It can be used to determine the dose to increase, decrease, suspend or maintain. Depending on the route of administration, the dose can be calculated according to body weight, body surface area or organ. The final dosage regimen will be considered by the attending physician in good medical practice to change the role of the drug. Determined by various factors, such as 163053.doc •67· 201242598 The specific activity of the agent; the nature and severity of the disease state; the patient's reactivity; the patient's age, condition, weight, sex and diet; and any infection Severity. Other factors that may be considered include time and frequency of administration, combination of drugs, sensitivity of response, and tolerance/response to therapy. Proficiency of the practitioner, especially based on the disclosed dose information and analysis, and in human clinical trials. Observed pharmacokinetic data 'The doses suitable for treatment can be routinely further improved without undue experimentation, including any of the formulations mentioned herein. Suitable doses can be used to determine the concentration of the agent in body fluids or other samples. The established analysis and dose response data were determined. The frequency of administration will be visualized. Depending on the pharmacokinetic parameters of the compound and the route of administration, the dosage and administration are adjusted to provide sufficient active fraction or to maintain the desired effect. Thus, the pharmaceutical composition can maintain the single dose, multiple doses, and Continuous infusion, sustained release sump or a combination of them. Short-acting pharmaceutical compositions (ie shorter half-life) can be administered once a day or more than once a day (eg twice, three times or four times a day). The drug may be administered every 3 to 4 days, every week, or every two weeks. Pumps such as subcutaneous, intraperitoneal or subdural pumps may be preferred for continuous infusion. Individuals that respond smoothly to the methods of the invention typically include medicine And veterinary subjects, including human patients. Other subjects to which the methods of the invention are applicable are tracing, dogs, large animals, birds (such as chickens), and the like. In general, any individual benefiting from a compound of formula A is suitable for administration of the methods of the invention. In some of the above embodiments, the patient has a cytogenetic character of del(17p)4del. In some of the above embodiments, the patient has lymphadenopathy. In some of the above embodiments, 'using Compound I, Η or H" will drop 163053.doc • 68· 201242598 The size of lymphatic lesions in low patients. In some of the above examples, the use of Compound I, Γ, II or II" reduces the size of lymphatic lesions after one treatment cycle In some of the above embodiments, the use of a compound "worker" or IΓ reduces the size of the lymphatic lesion by at least 10/〇 after one treatment cycle. In some of the above examples, the use of Compound I, I”, II or II" reduces the size of lymphatic lesions by at least 250/ after a treatment cycle. In some of the above examples, the use of Compound I, I ", η or η" will result in lymphoid glands after one treatment cycle. The size of the lesion is reduced by at least 30%. In some of the above-described embodiments, 'using Compound I, I, II, or II" reduces the size of lymphatic lesions by at least 40% after one treatment cycle. In some of the above examples, the use of Compound JJ", ΙΓ, reduces the size of lymphatic lesions by at least 50% after one treatment cycle. In some of the above embodiments, Compound 1, Ρ, II, or II" is used in a treatment cycle. The size of the lymphatic lesion can then be reduced by at least 75%. In one aspect, the invention provides a method of treating a condition comprising administering to a subject in need of such treatment, a compound of formula π or a pharmaceutically thereof thereof An acceptable salt and one or more therapeutic agents, wherein the condition is cancer. In one embodiment, the one or more therapeutic agents are proteasome inhibitors. One or more therapeutic agents may include bendamustine, rituximab Infliximab, orfarizumab and/or lenalidomide. In another embodiment, - or a plurality of therapeutic agents including bendamoxime / gamma, rituximab or both therapeutic agents In another embodiment, the one or more therapeutic agents include orfarizumab. In another embodiment, the one or more therapeutic agents include lenalidomide. The suitable dosage range of one or more therapeutic agents can vary. But in general, 16305 3.doc •69- 201242598 One or more therapeutic agents are administered beta in the range of 50 mg/m2 to uoo mg/m2. In one embodiment, bendamustine is administered in the range of 5 〇mg/m2 In another embodiment, the rituximab is administered in a range from 3 mg/m to 400 mg/m2. In another embodiment, the olfaxumab is in the range of 300 mg/m2 Administration of one or more therapeutic agents in combination with a compound of formula A for at least one cycle. In some embodiments, administration of one or more therapeutic agents in combination with a compound of formula A can last for at least 7 days. In other embodiments The administration of one or more therapeutic agents in combination with a compound of formula A can last for about 28 days. In some embodiments, the compound of formula A and the one or more therapeutic agents are each administered at least once during at least one cycle. The one or more therapeutic agents can be administered to the individual in the same or different period of time as the administration of the compound. In some embodiments, the one or more therapeutic agents are administered to the individual on at least the first and second days of at least one cycle. In other embodiments, one or more therapeutic agents are administered to the individual weekly. The assessment of the patient's clinical response can be measured after each cycle. Clinical results can be used to determine the increase, decrease, suspension or maintenance of the dose. In some of the above embodiments, the condition is a hematological malignancy. In a preferred embodiment, the condition is selected from the group consisting of multiple myeloma acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, B-cell lymphoma, diffuse large B-cell lymphoma, B cell ALL, T cell ALL, and Hodgkin's lymphoma. In a preferred embodiment, the compound substantially comprises the s-enantiomer. In a particular embodiment, the compound comprises at least 95 Å/〇 of the S·enantiomer. In some of the above examples, (4) synergistic benefits obtained by the compound and the therapeutic agent 163053.doc • 70· 201242598 are superior to those obtained without the combination of the compound and the therapeutic agent. The following examples are intended to illustrate and not to limit the invention. In the following examples, reference to "a compound of formula I" or "compound I" refers to the S-enantiomers shown herein, and the samples used in such examples exhibit 98.2% ee, as by Measured by HPLC method:

In addition, analysis of this compound revealed the following characteristics of the material: Test test results Appearance slightly grayish white powder 1H-NMR spectrum conformed to reference HPLC analysis 98.1% (based on anhydrous, solvent free) for palm purity (HPLC) 98.2% ee test results Burning residue 0.11% Infrared spectroscopy (FTIR) Spectra compliant with reference 13C-NMR Spectra of reference particle size analysis Median diameter: 11.3 μτη Water (Coulometric Karl Fischer) 0.56% Property or test Results C, Η, F, Ν element analysis % Expected value Experimental value %C 63.3 63.5 %H 4.4 4.4 %N 23.5 23.1 %F 4.5 4.5 163053.doc -71 - 201242598 Example 1 Inhibition of MM cell cell growth This example demonstrates Compound I inhibits cell growth stimulating effects of cytokines (IGF-1 and IL-6) on multiple myeloma (MM) cells. In the presence or absence of IL-6 or IGF-1, LB cells (bone marrow monocytic myeloma cell line) were incubated with the compound of formula I for 48 hours with control medium. The growth of MM cells was evaluated by measuring the absorbance of 3-(4,5-dimercaptothiazol-2-yl)-2,5-diphenyltetrasodium bromide (MTT; Chemicon International) dye. Inhibition. At the last 4 hours of culture for 48 hours, 10 pL of 5 mg/mL MTT was added to each well to pulse-stimulate the cells, followed by the addition of 100 μί of isopropanol containing 0.04 N of HC1. Absorbance was measured at 570/630 nm using a spectrophotometer (Molecular Devices). An overview of the results is shown in Figure 1. Exposure to 0.625 μΜ-2.5 μΜ of Compound I inhibited MM cell growth even in the presence of cell growth stimulating cytokines. Example 2 Effect of BMSC on Cytotoxicity This example demonstrates that bone marrow stromal cells (BMSC) do not prevent Compound I-induced LB cell cytotoxicity. LB cells were incubated with the control broth in the presence or absence of BMSC and incubated with the compound of formula I for 48 hours. Cell proliferation was assessed using [3Η]-thymidine uptake assay. All data represent the average of three replicate experiments (soil SD). An overview of the results is shown in Figure 2. After exposure to 0.625 μΜ-I0 μΜ Compound I, LB cell growth was reduced even in the presence of BMSC. Example 3 Effect of Compounds on Apoptosis of CLL Cells 163053.doc • 72- 201242598 This example demonstrates that the compound of formula i induces apoptosis in chronic lymphocytic leukemia (CLL) cells of patients. Peripheral blood was obtained from patients with B-CLL via the CLL Research Consortium from Ohio State University. Primary CD19 positive cells were isolated using Rosette-Sep (StemCell Technologies). The cells were supplemented with 10% heat-inactivated fetal bovine serum, 2 mmol/L L-glutamic acid and penicillin (100 units/ml) / lentin at 37 ° C, 5% CO 2 and high humidity. (Streptomycin) (100 pg/mL; Invitrogen) maintained in RPMI 1640 (Invitrogen). After 96 hours of incubation with the compound of Formula I or the medium, 5χ105 cells were washed with PBS and then resuspended in 2 μΐ^ Annexin V-FITC stock solution (BioWhittaker, Inc) and 10 μΐ^ 20 pg /mL Pl (Sigma) binding buffer (10 mmol/L HEPES/NaOH (pH 7.4), 150 mmol/L NaCl, 5 mmol/L KC, 1 mmol/L MgCl2, 1.8 mmol/L CaCl2). After incubation for 10 minutes at room temperature in the dark, the samples were quantified by flow cytometry on a FACScanTM (Becton Dickinson). Treatment of CLL patient cells with Compound I caused apoptosis and the results appeared to be dose dependent, as seen in Figure 3. Compound I induced apoptosis in CLL cells from patients with poor prognosis, as indicated by the data in Figure 19. Compound I was also effective in inducing cell death in CLL cells in refractory/relapsed patients, as shown in Figure 20. Example 4 Effect of Compounds on ALL Cell Lines 163053.doc •73- 201242598 This example demonstrates that compounds of formula I cause a decrease in Akt acidification and cell proliferation in leukemia-ALL and B-ALL (acute lymphoblastic leukemia) leukemia cell lines. Small, accompanied by cell death. Cell lines were analyzed for viability using AlamarBlue assay (Invitrogen). Cells of 100 pL in volume (1χ106 per well) were placed in a 96-well flat pan and a compound of formula I (100 pL per well, 2 χ final concentration) or medium alone was added to each plate. All experiments were performed in 4 replicates. The cells were incubated for a fixed time (48 hours). After the incubation, 10 gL of AlamarBlue® was added to each well. The cells were incubated for 4 hours and the optical density at 530-560 nm was obtained using a SpectraMax® M5 disk reader 2001. Cell viability is expressed as the percentage of absorption between treated cells/control samples. These results are summarized in the table shown in Figure 4. Exposure to Compound I caused a substantial decrease in cell viability and decreased Akt phosphorylation in various ALL cell lines. Example 5 Effect of Compounds on ALL Cell Cycle This example demonstrates that treatment of acute lymphoblastic leukemia (ALL) cell line CCRF-SB with a compound of formula I causes G0/G1 cell cycle arrest. Representative luminescence activated cell sorting (FACS) analysis was performed on CCRF-SB cells stained with propidium iodide under normal growth conditions and CCRF-SB cells grown in the presence of a compound of formula I. The average percentage of Go-G!, S, and G2-M phase cells is calculated in the table below the histogram. The results are shown in Figure 5. Example 6 inhibits proliferation of breast cancer cells

This example demonstrates that the compounds of formula I inhibit the proliferation of breast cancer cell lines. T47D 163053.doc -74- 201242598 and HS-5 78T cell lines were grown in serum in the presence of a compound of formula I at the indicated concentrations. Proliferation was measured in three replicate wells of a 96-well plate by AlamarBlue® analysis (Invitrogen). The results of the proliferation assay are expressed as mean cell percentage values and are shown in Figure 6. Example 7 Inhibition of Proliferation of Ovarian Cancer Cell Lines This example demonstrates that compounds of Formula I inhibit proliferation of ovarian cancer cell lines. IGROV-1 and OVCAR-3 cell lines were grown in serum in the presence of a concentration of a compound of formula I. Proliferation was measured in three replicate wells of a 96 well plate by AlamarBlue analysis (Invitrogen). The results of the proliferation assay are expressed as mean cell percentage values and are shown in Figure 7. Example 8 Reduction of Akt Phosphorylation This example demonstrates that a compound of formula I reduces constitutive Akt squamation in a blood tumor cell line exhibiting constitutive Akt phosphorylation. Constitutive Akt phosphorylation of a large group of leukemia and lymphoma cell lines was assessed. These cell lines represent B-lymphoma, T-lymphoma, ALL, malignant histiocytosis, DLBCL and AML. Cell lines showing serum-independent Akt phosphorylation were treated with the compound of formula I for 2 hours. Thereafter, the cells were lysed, size-fractioned, and immunoblot analysis was performed using an antibody against acidified Akt (Ser473). The results are shown in Figure 8. After exposure to Compound I, all cell lines achieved Akt (Ser473) reduction. Example 9 Compound I is effective in DLBCL 163053.doc -75- 201242598 This example demonstrates that Compound I blocks PI3K signaling and induces apoptosis in diffuse large B-cell lymphoma cells. ΡΙΙΟδ is expressed in the DLBCL cell line as shown in Figure 26A. Figure 26 shows that exposure to Compound I reduces the content of ρΑΚΤ in several DLBCL cell lines. Example 10 Induction of Apoptosis in Breast Cancer Cells This example demonstrates that a compound of formula I induces apoptosis in a breast cancer cell line. HS-578 Τ, T47D and MCF7 cells were treated with the compound of formula I or the corresponding DMSO concentration for 24 hours. The percentage of apoptotic cells was determined by Annexin V-FITC/7AAD staining. On the bottom left side, live cells (Annexin V-FITC/PI negative); bottom right, early apoptotic cells (only annexin V-FITC positive); top right, metaphase/late apoptotic cells (Annexin V) -FITC/7AAD double positive); and top left, late apoptotic/necrotic cells (only 7AAD positive). In addition to the bottom left quadrant (live cells), the percentage of cells in each quadrant is indicated. An experiment representing three different experiments with similar results is shown in Figure 1〇. Example 11 Steady Blood Content on Day 7 in Healthy Volunteers This example provides information on the concentration of the compound of formula I in the blood of healthy human subjects on day 7. The concentration during the 12 hour period was monitored after oral administration of 5 〇, 1 〇〇 or 200 mg BID of the compound of formula I on the 7th day of the study. Figure 11 tracks the plasma concentration of the drug during the 12 hour period from the time of administration. All doses reached the maximum drug concentration within two hours. Administration of this compound at 5 〇, 1 〇〇 or 2 〇〇 BID causes the concentration level to exceed the EC5 〇 concentration in the basophilic blood cells for at least 12 hours. 163053.doc •76- 201242598 In addition, a single dose study was conducted in which 17-400 mg of the compound of formula I was administered to healthy volunteers. The concentration of the compound in the blood for 24 hours from the administration of the drug was measured and the results are shown in Fig. 24A. At about 6 hours, Compound I has a concentration in the blood of at least about 100 nM for all administered doses. At about 12 hours, the concentration of Compound I in the blood exceeds 50 nM for doses of 50 mg and above. The maximum concentration of Compound I in the blood was achieved within 2 hours of administration. In another experiment, the mean Compound I concentration was measured on day 7 of 50 mg BID given to healthy volunteers (N=6). The mean lowest concentration is higher than the EC50 for ΡΙ3Κδ and the average peak concentration is lower than the EC50 for ΡΙ3Κγ, as determined in the whole blood basophil hematopoietic assay, Figure 24Β. This example demonstrates that the concentration range of Compound I administered at 50 mg BID is above the ED5〇 Κ3Κδ basophilic hematopoietic level in whole blood, but below the minimum ED5〇 ΡΙ3Κγ basophil activation level for at least 12 hours. Table 1 below provides an overview of the individuals in the study in which a single dose (SD) or multiple doses (MD) of a compound of formula I is administered to individuals in varying amounts. The "η" value refers to the number of individuals in each group. Table 1 Group Protocol Compound 1 Placebo 1 (η=8) SD 17 mg (n=6) Placebo (n=2) 2 (η=8) SD 50 mg (n=6) Placebo (n=2) 3 (η=8) SD 125 mg (n=6) placebo (n=2) 4 (η=8) SD 250 mg (n=6) placebo (n=2) 5(η=8) SD 400 mg (n=6) placebo (n=2) 6 (η=8) MD 50 mg BID x 7 days (n=6) placebo BID x 7 days (n=2) 7 (η=8) MD 100 mg BID x 7 days (n=6) placebo BID x 7 days (n=2) 8 (η=8) MD 200 mg BID x 7 days (n=6) placebo BID x 7 days (n=2) 163053 .doc •77- 201242598 Example 12 Effect on lesions in patients with mantle cell lymphoma This example provides information on lesions in patients with mantle cell lymphoma after one cycle (28 days) of treatment with a compound of formula I Area information. The area of 6 lesions was measured before treatment and after the treatment period. The response to a 28-day oral administration of 50 mg BID of the compound of formula I caused a reduction in the area of the lesion compared to the area before treatment and a 44% reduction in tumor burden. The results are summarized in the bar chart seen in Figure 12. Example 13 Response of CLL patients to treatment This example provides information on the concentration of absolute lymphocyte counts (ALC) in the blood of CLL patients after one cycle (28 days) of oral administration of the compound of formula j. Blood ALC concentrations were measured during the 4 weeks after completion of one treatment cycle. It was observed that lymphocytosis was reduced by 55% due to treatment and lymphatic lesions were reduced by 38% due to treatment. A significant decrease in ALC concentration was observed between week 1 and week 2, Figure 13. Example 14 Comparing Lymphoma Patients with Healthy Volunteers This example provides data comparing the concentrations of compounds in lymphoma patients with normal healthy volunteers. On day 28, a compound of 5 mg mg Bm was orally administered to a patient with mantle cell lymphoma, and the concentration of the compound in the blood was measured during 6 hours after administration. The concentration of $ and 1 mg administered orally in normal healthy volunteers on the 7th day of dosing was also observed. The results are summarized in Figure 14. Thus, the compound does not accumulate excessively during the treatment cycle, and the 163053.doc -78- 201242598 does not become responsive to metabolic enhancement during the treatment cycle. Example 15 Activity of Compound I against various kinases This example demonstrates the IC5Q profile of Compound I for each class of kinases, as outlined in Table 2. Although particularly active against ρΐ ι0δ, Compound I is also active against ρΠΟγ and is even active enough to be therapeutically suitable for combating ρΐ 1〇β ' at non-toxic doses because the compound has been shown to have a higher NOAEL rating; -V-type phosphoinositide kinase exhibits little activity. Thus, despite the delta selectivity, the activity exhibited by the compound on ρ1 ι〇γ may be sufficient for clinical applicability', ie, effective for cancers that rely on ρ110γ for signal transduction, because plasma levels are higher than effective inhibition of pi 1 〇 The dose of gamma, while still being selective with respect to other isoforms, specifically alpha isoforms. Table 2

Class 1 PI3K, IC50(nM) Class II ΡΙ3Κ, IC5〇(nM) Class III PI3K * ICs〇(nM) Class IV PI3K, !C5〇(nM) Other acid inositol kinase compound pl 10α ρΙΙΟβ pllOS ρΙΙΟγ αΐβ hVPS34 DNA- PK mTOR ΡΙΡ5Κα ΡΙΡ5Κβ I 435 128 1 14 >103 978 6,729 >103 >ιο3 1 >103 NVP-BEZ- 235 Novartis 19 293 63 267 3 6 1 2 ND

InvitroGen Adapta Analysis * ND = Not Detected Example 16 Compound I has no detachment activity in whole-stranded histone kinase screening. This example demonstrates that Compound I has a very high level of histone kinase screening in 163053.doc -79 - 201242598 Little or no off-target activity. Using the Ambit KINOMEscanTM, a genome-wide screening of more than 350 protein kinases at 10 μΜ failed to detect any activity. Examples of some of the kinases in the screening are shown in Table 3 below. Table 3 Examples of related kinases in screening ABL FGFR1 JAK1 P38MAPK S6K AKT VEGFR1 JAK2 PDGFR SLK ALK FLT3 JNK1 PIM SRC BLK FRK KIT PKA SYK BRAF FYN LCK PKC TAK BTK HCK LYN PLK TIE CDK HER2 MAPK RAF TRK CSF1R ICK MEK RET TYK EGFR IGF1-R MET ROCK YES EPH ITK MLK ROS ZAP70 Example 17 Compound I selectivity for Ρ1105 This example demonstrates that Compound I is selective for ρΐ 1〇δ, as measured in isoform-specific cell-based assays. .

Swiss-3T3 fibroblasts and RAW-264 were seeded on 96-well tissue culture plates and allowed to reach at least 90% confluence. The cells were starved and treated with vehicle or serial dilutions of Compound I for 2 hours and stimulated with PDGF or C5a, respectively. Akt phosphorylation and total AKT were detected by ELISA. Purified B cells were treated with vehicle or serial dilutions of Compound I for 30 minutes at room temperature, followed by the addition of purified goat anti-human IgM. The results are expressed as relative [3H] thymidine incorporation induced by IgM crosslinking. 163053.doc • 80 - 201242598 Table 4 ΡΙ3Κα EC5〇(nM) ___ ΡΙ3Κδ ECs〇(nM) ΡΙ3Κγ EC5〇(riM) fibroblast cell line primary B cell monocyte cell line PDGF-induced ρΑΚΤ BCR-mediated proliferation C5a-induced PAKT > 20,000 6 3,894 (η=12) (n=6) (η=11) Example 18 Performance of pll〇5 in leukemia and lymphoma cell lines This example demonstrates that PI3K ρΙΙΟδ is highly expressed in a wide range. In leukemia and lymphoma cell lines. ΡΙ3Κ ρ 110δ promotes proliferation and survival of a wide range of leukemia and lymphoma cell lines. The cell types studied were MCL, DLBCL, AML, ALL and CML. The expressions of PI3K ρ 110α, ρ 110β, ρΙΙΟγ, and ρΙΙΟδ in a group of lymphoma and leukemia cell lines are shown in Fig. 15. Proteins from 106 cells were separated by sdS-PAGE and analyzed using Western blotting using antibodies specific for α, β'γ and δ isoforms. Purified recombinant pi 10 protein was used as a control. Anti-actin antibodies were used to evaluate samples of equal loading. Ρΐ 1〇δ is consistently expressed in high amounts, while other pllO isoforms are highly variable. It is known that ΡΙ3Κ ρΐι〇δ is uniformly expressed in AML· cells of patients, as discussed by Sujobert, et al., B/ood 2005 106(3), 1063-1066. Example 19 Inhibitory effect of Compound I on pllOS Example 19 shows that Compound I inhibits Ρ11 〇δ to block leukemia and lymphoma 163053.doc • 81 · 201242598 Cell line with PI3K signaling with constitutive pathway activation. In leukemia and lymphoma cell lines, the PI3K pathway is often abnormally regulated. It was found that 48% of the cell lines, or 13/27 of the cell lines, had a constitutive p-AKT. Furthermore, PI3K path activation is dependent on ρΙΙΟδ. Compound I was found to inhibit constitutive ΑΚΤ phosphorylation in a 13/13 cell line. The PAGE results of Fig. 9 demonstrate that constitutive ΑΚΤ phosphorylation is inhibited by the presence of Compound I in each of 11 cell lines including B cells and T cell lymphoma. The cells were incubated with 10 μM of Compound I for 2 hours. The cell lysate was subjected to SDS-PAGE and transferred to a PDVF membrane and probed with an appropriate antibody. Compound I was found to inhibit constitutive ΑΚΤ phosphorylation in a 11/11 cell line. Additional cell lines of T-ALL and B-ALL cell lines are shown in Figure 27. The Akt and S6 phosphorylation reduction after exposure to a range of concentrations of Compound 1 (0.1 μΜ to 10 μΜ) was quantified by densitometry, expressed as percent change, Figure 28. Example 20 Compound I inhibits proliferation of leukemia cell lines and induces apoptosis. Example 20 demonstrates that Compound I inhibits proliferation and induces apoptosis in leukemia cell lines. Figures 16A-B show that treatment with Compound I for 24 hours reduced cell viability in a dose-dependent manner. Proliferation assays (AlamarBlue®) were performed on ALL cell lines grown in the presence of 10% FBS serum at 24 hours and were measured. Proliferation was measured in three replicate wells of a 96-well plate. Inhibition of PI3K signaling by Compound I causes cell cycle progression and/or cell death. In each of the six leukemia cell lines, Compound I at a concentration of 10 micromolar reduced the survival rate by 40-163053.doc -82·201242598 50%, and Figure 16A induces apoptosis of Compound I. Cells were treated with DMSO (vehicle), 1 μM or 10 μM Compound I for 24 hours. The percentage of apoptotic cells was determined by Annexin V-FITC/7AAD staining. Representing different experiments with similar results - an experiment is shown in Figure 16Β. Example 21 Performance of pllO 5 in CLL cells This example demonstrates that PI3K ρΙΙΟδ and ρΙΙΟδ isoforms behave in patient CLL cells. The signal transduction pathway mediated by ΡΙ3Κ has been implicated in CLL. These pathways play a role in cell proliferation, arrest of apoptosis, and cell migration. An attempt was made to determine the presence of a ΡΙ3Κ isoform in a patient's CLL cells. The demographic status of CLL patients is summarized in Table 5 below. Table 5 Demographic status of CLL patients (total (7V=24)) I) cytogenetic abnormalities 13ql4.3 58% llq22.3 33% 17ρ13·1 20% 12th trisomy 12% II) Treatment of striated Labin is difficult to cure 29% Unknown 54% II) IgVH state mutation 33% Unmuted 33% Unknown 33% 163053.doc -83· 201242598 Figure 17A-D PAGE image comparison ρΐι〇α, ρΐι〇δ, ρΙΙΟβ and ρΙΙΟγ ο Performance in CLL cells of patients with sputum. ρΙΙΟδ and ρΙΙΟγ are expressed in each patient compared to other ΡΙ3Κ isoforms. Example 22

Compound I induces caspase 3 and PARP. This example demonstrates that Compound I induces the splitting of caspase 3 and PARP. Figure 18A-B shows the results of caspase 3 and PARP (poly(ADP) ribose polymerase) cleavage in the presence of 1 μΜ, 10 μΜ of Compound I or 25 μM of LY294002. Other experiments provide evidence that Compound I induces caspase 2 and PARP cleavage. The cells were incubated with Compound I or vehicle alone for 24 hours. Thereafter, the cells were lysed and size fractionated and immunoblot analysis was performed with antibodies against FLIP, Figure 29. In addition, whole cell lysates were added to a MDS (Meso Scale Diagnostics) multi-point 96-well 4-point plate coated with total caspase-3, cleaved caspase-3, split PARP and BSA. Proteins were detected and quantified using antibodies labeled with SULFO-TAG reagent. A dose-dependent reaction of split caspase 3 and PARP was achieved by exposure to 5 μΜ or 10 μΜ of Compound I. Example 23 Compound I blocks P13K signaling This example demonstrates that Compound I blocks PI3K signaling in AML cells in patients. ΡΙ3Κδ is involved in signal transduction in AML patient cells. Figure 21 shows the results of carbonation of Akt in the absence or presence of 0.1, 1.0, 10 μM of Compound I. This demonstrates that Compound I reduces phosphorylation of Akt in patients with AML 163053.doc • 84· 201242598. Example 24 Measurement of PI3K signaling in basophilic blood cells based on whole blood measurements This example shows the use of flow cytometry to measure whole blood analysis of PI3K signaling in basophilic blood cells by inducing CD63 surface appearance. Inhibition of PI3K signaling in basophilic blood cells allows Compound I to be a pharmacodynamic marker. PI3K signaling was monitored by CD63 surface performance. In particular, ρΙΙΟδ mediates FCsR1 signaling and ρΙΙΟγ mediates fMLP receptor signaling. Flow cytometry analysis of PI3K-mediated CD63 expression on basophilic blood cells involves the following sequential steps: 1. Collection of peripheral blood 2. Basophilic hematocytosis (fMLP or anti-FCsRIMab) 3. Labeling basophilic Blood cells (anti-CCR3-FITC and anti-CD63-PE) 4. Dissolve and fix cells 5. Analyze by flow cytometry Figure 22A-C compares A) no stimulation, B) stimulation with anti-FCsRl or C) with fMLP The result of the stimulus. Figure 23 shows that Compound I is particularly active when p 110δ-mediated signaling is most important, but is also relatively active when using ρΙΙΟγ: for the ρΙΙΟδ test, it achieves a 50% reduction in SD63 performance at 1 μΜ, and for ρΙΙΟγ Test, which achieved a 50% reduction in SD63 performance at approximately 10 μΜ. The Flow2 CAST® kit was used to measure basophilic blood cell activation in human whole blood. Whole blood samples were treated with vehicle or serial dilutions of Compound I, followed by activation of basophilic blood cells with anti-FcsRI mAb or fMLP. Cells were stained with anti-human CD63-163053.doc •85- 201242598 FITC in combination with anti-human CCR3-PE mAb. The percentage of CD63 positive cells in the basophilic hematocrit population in the different treatment groups was determined and corrected for vehicle control. Example 25 Compound I attenuates lymphadenopathy in CLL patients This example provides evidence of size reduction in large lymph node lesions in CLL patients with del[17p]. Patients with del (17p) had an axillary lymphadenopathy, which was imaged by computed tomography (CT) to provide a baseline measurement of 5.9 cm x 4.1 cm, Figure 4〇a. After one cycle of treatment with Compound I, the lymphadenopathy was reduced to a size of 3.8 x 1.8 cm, Figure 40B. One cycle of treatment was continued for oral administration with Compound I of 200 mg BID or 350 mg BID for 28 days. Example 26 Limited Effect of Compound I on a Glucose and Insulin Content of This Example This example demonstrates that treatment with a compound has little or no effect on glucose and insulin levels. Compound j was administered to the individual twice daily at a rate of 5 〇 2 〇〇 mg over a period of up to 1 day. Blood glucose and insulin concentrations were measured over time and compared to placebo results as shown in Figures 25A-B. The blood glucose concentration remained stable even after 10 days of treatment with the highest dose of the compound. In use of compounds! After 7 days of treatment, the insulin content remained within the normal range. This demonstrates that the compound quinone has little or no effect on glucose and insulin levels. • Example 27 Materials and Methods 163053.doc • 86· 201242598 This example provides information on the materials and methods for conducting the experiments described in Examples 28-35 regarding the use of Compound I for the treatment of multiple bone tumors. Materials ρΐ 10δ Inhibitors Compound I and Compound II were supplied by Calistoga Pharmaceuticals (Seattle, WA). The S enantiomers in the samples of compounds I and II used exceeded 95%. Compound I was dissolved in dimethyl sulfoxide at 10 mM and stored at -20 °C for in vitro studies. Compound II was dissolved at 1 °/. Carboxymethylcellulose (CMC) / 0.5% Tween 80 (Tween 80) and stored at 4 ° C for in vivo studies. Recombinant human ΡΙΙΟα, ΡΙΙΟβ, ΡΙΙΟγ, and Ρ110δ were reconstituted with sterile phosphate buffered saline (PBS) containing 0.1% BSA. Gentazole is supplied by Millennium Pharmaceuticals (Cambridge, MA). 3-methyl gland ° ticket was purchased from Sigma-Aldrich (St. Louis, MO). Cell culture

Dex Sensitivity (MM.1S) and Resistance (MM.1R) human MM cell lines were kindly provided by Dr. Steven Rosen (Northwestern University, Chicago, IL). H929, RPMI8226 and U266 human MM cell lines were obtained from the American Type Culture Collection (Manassas, VA). The melphalan-resistant RPMI-LR5 and cranberry (Dox) resistant RPMI-Dox40 cell lines were kindly provided by Dr. William Dalton (Lee Moffitt Cancer Center, Tampa, FL). OPM1 public cell leukemia cells were provided by Dr. Edward Thompson (University of Texas Medical Branch, Galveston). The IL-6-dependent human MM cell line INA-6 was supplied by Dr. Renate Burger (University of Kiel, Kiel, Germany) 163053.doc -87-201242598. LB human MM cell strains are produced in the laboratory. Phenotypic analysis revealed acellular genetic abnormalities. Phenotypic analysis is shown in Table 6. The CD appearance profile of LB cell lines was determined by flow cytometric analysis. Table 6 LB expression CD marker Performance % CD3 5.5% CD19 61.7% CD20 97.2% CD38 54.1% CD40 96.8% CD49e 5.9% CD70 98.0% CD138 96.3% All MM cell lines were cultured in RPMI1640 medium. Bone marrow stromal cells (BMSC) improved in Dulbecco containing 15% fetal calf serum, 2 mM L-glutamic acid (Life Technologies), 100 U/mL penicillin and 100 pg/mL streptomycin (Life Technologies) Cultured in Dulbecco's modification of Eagle's medium (DMEM) (Sigma). Blood samples collected from healthy volunteers were processed to obtain peripheral blood mononuclear cells (PBMNC). After obtaining the informed consent form according to the Declaration of Helsinki and approved by the Institutional Review Board of the Dana-Farber Cancer Institute (Boston, MA), from the BM sample Patient MM and BM cells were obtained. BM monocytes were isolated using Ficoll-PaqueTM density sedimentation and positively selected by separation of microbeads (Miltenyi Biotec, Auburn, CA) with anti-CD138 magnetic activation cells 163053.doc -88 - 201242598

Plasma cells were purified (>95% CD138+). Tumor cells were also purified from BM from MM patients using the RosetteSep Negative Selection System (StemCell Technologies, Vancouver, BC, Canada). Growth inhibition assay was assessed by measuring the absorbance of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrasodium bromide (MTT, Chemicon International, Temecula, CA). Growth inhibition of compound I on the growth of MM cell line, PBMC and BMSC" Effect of compound I on the growth of paracrine MM cells in BM MM cells (2χ104 cells/well) were coated with BMSC in the presence or absence of drug. Incubate for 48 hours in a 96-well plate (Costar, Cambridge, ΜΑ). DNA synthesis was measured by [3H]-brain (Perkin-Elmer, Boston, MA) uptake, with [3H]-thymidine (0.5 μCi/well) added during the last 8 hours of 48 hour culture. All experiments were performed in four replicates. Short-term blockade of P1105 expression using the cell line Nucleofector kit V (Amaxa Biosystems Gaitherburg, MD) with siRNA ON-TARGET plus SMART pool ΡΙΙΟδ or non-specific control duplex (Dharmacon Lafayette, Co) for transient transfection of INA-6 cells and LB cells. Immunofluorescence live MM cells (2.5 χ 104 cells) were assembled on glass slides by centrifugation at 500 rpm for 5 minutes using a cell centrifugation system (Thermo Shandon, Pittsburgh, PA). The cells were fixed in cold absolute acetone and methanol for 10 minutes. After fixation, the cells were washed in phosphate buffered saline (PBS) and then blocked with 163053.doc -89 - 201242598 PBS containing 5% FBS for 60 minutes. The slides were then incubated with anti-CD 138 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for 24 hours at 4 ° C, washed in PBS, and incubated with goat anti-mouse IgG for 1 hour at 4 ° C, and Analysis was performed using Nikon E800 fluorescence microscopy. The acid vesicle organelle (AVO) was measured and quantified using acridine orange staining values. Autophagy is characterized by chelation of cytoplasmic proteins and formation of AVO to detect and quantify AVO in cells treated with compound I or 3MA, and in vivo staining with acridine orange at a final concentration of 1 pg/ml for 15 minutes. The sample was examined under a fluorescent microscope. Angiogenesis assay The anti-angiogenic activity of Compound I was determined using an in vitro angiogenesis assay kit (Chemicon, Temecula, CA). HUVEC and endothelial growth medium were obtained from Lonza (Walkersville, MD, USA). HUVEC was incubated with Compound I on a polymeric matrix gel at 37 °C. After 8 hours, angiogenesis was assessed using Leika DM IL microscopy (Leica Microsystems, Wetzlar, Germany) and analyzed with IM50 software (Leica Microsystems Imaging Solutions, Cambridge, UK). HUVEC cell migration and rearrangement were observed and the number of branch points was counted. Western MM cells were cultured with or without Compound I; collected; washed; and used radioimmunoassay (RIPA) buffer, 2 mM Na3 V04, 5 mM NaF, 1 mM Methylsulfonate (5 mg /ml) dissolves. Whole cell lysate was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) at 163053.doc •90· 201242598, transferred to pure nitrocellulose membrane (Bio-Rad Laboratories, Hercules, CA) And using anti-AKT, phosphorylated (p)-AKT (Ser473, Thr308), ERKl/2, P-ERKl/2, P-PDKl, STAT'P-STAT, P-FKRHL, P-70S6K, LC3 and ΡΙ3Κ/ρ110α Ab (Cell Signaling Danvers, MA); anti-ρΙΙΟβ, ΡΙ3Κ/ρ110δ, 3-glyceraldehyde dehydrogenase (GAPDH), α-tubulin and actin Ab (Santa Cruz Biotechnology, CA); and anti-ρΐ 10γ Ab (Alexis, San Diego, CA) and anti-LC3 Ab (Abgent, San Diego, CA) for immunological dot analysis

ELISA The cytokine secretion by human BMSC co-cultured with MM cells was assessed by ELISA. BMSCs were incubated with different concentrations of Compound I in 96-well plates with or without INA-6 cells. After 48 hours, the supernatant was collected and stored at -80 °C. Cytokines were measured using a Duo set ELISA chromogenic kit (R&D Systems, Minneapolis, MN). All measurements were performed in three replicates. Human cytokine arrays were evaluated using the proteome Profiler antibody array set A (R&D Systems, Minneapolis, MN) according to the manufacturer's instructions for cytokine content in culture supernatants, and supernatants from BMSC co-cultures. The membrane of the Ab array of 37 cytokines was incubated for 4 hours. Mouse MM xenograft model of human MM CB17 SCID mice (48-54 days old) were purchased from Charles River Laboratories (Wilmington, ΜΑ). All animal studies were performed according to the protocol approved by the Animal Ethics Committee of the Dana-Farber Cancer Institute, Dana-Farber Cancer Research Institute 163053.doc -91 - 201242598. 3 x 106 LB cells contained in 100 μί RPMI-1640 were subcutaneously inoculated into the right abdomen of the mice. When the tumor was palpable, the mice were assigned to the treatment group and received 10 mg/kg or 30 mg/kg twice daily; and 7 of the mice in the control group received vehicle alone. The caliper gauge was measured every other day for the longest vertical tumor diameter to estimate the tumor volume using the formula representing the 3D volume of the ellipse: 4/3 χ (width/2) 2 χ (length/2). Animals were sacrificed when the tumor reached 2 cm or the mouse appeared to be dying. The survival was assessed until death on the first day of treatment. Tumor growth was assessed using caliper measurements from the first day of treatment until the day of the start of the sacrifice, which was day 12 for the control group and days 17 and 19 for the treatment group. Capture images with the Canon IXY Digital 700 camera. Ex vivo analysis values for tumor images were captured at 40 u/0.60 (Leica, Heidelberg, Germany) using a LEICA DM IL microscope and a LEICA DFC300 FX camera. Human fetal bone grafts were implanted into CB17 SCID mice (SCID-hu). Four weeks after bone implantation, 2.5 χ 106 INA-6 cells were injected directly into the human sacral cavity in the graft in a final volume of 100 μΐ RPMI-1640 medium. The increase in the content of soluble human IL-6 receptor (shuIL-6R) derived from ΙΝΑ-6 cells was used as an indicator of MM cell growth and disease burden in SCID-hu mice. Mice produced a measurable serum shuIL-6R about 4 weeks after INA-6 cell injection, and then received 10 or 30 mg/kg of drug or a separate vehicle daily for 7 weeks. Blood samples were collected and their shuIL-6R content was assessed using enzyme-linked immunosorbent assay (ELISA, R&D Systems. Minneapolis MN). Statistical Analysis 163053.doc -92- 201242598 Statistical significance was determined by Dunn's multiple comparison test. The minimum significant level is p<〇.〇5. Survival was assessed using a Kaplan-Meier curve and log rank analysis. The combined effect of Compound I and bortezomib was analyzed by isobologram analysis using the CalcuSyn software program (Biosoft, Ferguson, MO); the combined index (CI) < 0.7 indicates a synergistic effect. Example 28 Expression of pll〇5 in MM cells This example demonstrates that ρΐ 10δ is highly expressed in patient MM cells. To evaluate the ΡΪ3Κ/ρ110 performance' using Abs for recombinant human ρΐ3Κ/ρ110α, ρΙΙΟβ, ρΙΙΟγ, and ρΙΙΟδ proteins, these Abs have specific immunoreactivity for this equivalent work isoform. Evaluation of 11 MM cell lines (MM. 1 S, OPM1, OPM2, RPMI8226, DOX40, LR5, MM.1R, U266, INA-6, H929 and LB) and the expression of ρΙΙΟδ in 24 patient MM samples and immunoblotting Points are shown in Figures 30A and 30B. Figure 30A shows the expression of ρΙΙΟα, ρΙΙΟβ, ρΙΙΟγ, and ρι10δ in a sputum cell line detected by immunoblotting using a specific antibody. Anti-[alpha]-tubulin MAb served as a loading control. The anti-Ρ110δ Ab was used to detect pii in patient MM cells by immunoblotting analysis (Fig. 3〇B) e Anti-GAPDH MAb served as a loading control. ΙΝΑ-6 and LB cells strongly expressed ρΙΙΟδ, while MM.1S, OPM1, MM.1R, Dox40, U266 or H929 lacked ρΙΙΟδ expression (Fig. 30A). Confirmation of ρΙΙΟδ in MM.1S and LB cells by immunofluorescence analysis (Fig. 30C) » Human recombinant ριι〇α, ρι 1〇β, Ρ11〇γ, ρι 1〇δ protein 163053.doc •93- 201242598 The SDS sample buffer was heated for 3 minutes and then loaded onto the gel. (10-20 ^ per lane) Recombinant human ΡΙΙΟα, ΡΙΙΟβ, ΡΙΙΟγ ' Ρ110δ eggs were detected by immunoblotting analysis. Using ρι1〇δ specificity FITC-conjugated secondary antibody measurements mm 1S and lb cells in LB cells ΡΙΙΟ § ΡΙΙΟ ΡΙΙΟ δ δ ΡΙΙΟ 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 染 核酸 核酸Western blots reveal that there is no correlation between ρΙΙΟδ performance and other isoforms (α, β, and γ). It is important that all patient MM cells also exhibit ρΙΙΟδ (Fig. 30B). Example 29 Cytotoxicity of Compound I against Sputum Cells This example demonstrates that Compound I is selective cytotoxic against cells having ρΐ 10δ. In detail, 'Compound I strongly induces cytotoxicity in ρ11〇δ-positive sputum cells and in primary patient sputum cells, and is not cytotoxic in peripheral blood mononuclear cells from healthy donors, indicating a favorable therapeutic index. The ρ 11 0δ gene showed an inhibitory effect on the growth of sputum cells. LB and ΙΝΑ-6 cells were transfected with Ρ11〇δ siRNA (Si) or control siRNA (mimetic). After 24 hours, the expression of pi 1 〇δ was determined by Western blot analysis, see Fig. 31Α. ΙΝΑ-6 cells were transfected with ρΐ 10δ siRNA or control siRNA' and cultured for 72 hours. Cell growth was assessed by MTT assay, see Figure 31B. Data indicate the mean SD of the three replicate cultures, expressed as a multiple of the control. Transfection with ρΐ 10δ siRNA instead of mock siRNA down-regulated ρΙΙΟδ and inhibited MM cell growth at 72 hours (Figures 31A and 31B). The growth inhibitory effect of ρΐ 10δ-specific small molecule inhibitor Compound I on MM cell line, 163053.doc •94· 201242598 PBMC and patient MM cells was evaluated. Compound I induced cytotoxicity against LB and INA-6 MM cells (ρΙΙΟδ-positive) in a dose- and time-dependent manner; on the contrary, it was noted that cytotoxicity was minimal in the ρ110δ-negative cell line (Fig. 3 1C). Figure 3 1C legend: LB (d), INA-6 (A), rpmI 8226 (〇), 〇 PM2 (〇), Η 929 (·), U266», rPMI-LR5 (A) and ΟΡΜ 1 (·) ΜΜ Cells were incubated with or without Compound I for 48 hours. Importantly, Compound I also induced cytotoxicity against MM cells in patients (Fig. 31D) and was not cytotoxic in PBMC from 4 healthy volunteers at concentrations up to 2 〇 μΜ (Fig. 3丨Ε). The patient's sputum cells were cultured with Compound I for 48 hours by negative selection. Peripheral blood mononuclear cells isolated from healthy donors were incubated with Compound I for 72 hours. The data represents the mean value of SD survival rate as assessed by ΜΤΤ analysis of three replicate cultures as a percentage of untreated controls. These results strongly indicate that sensitivity to Compound I correlates with ρι 1〇5 performance and indicates a favorable therapeutic window. To determine whether Compound I induced cytotoxicity via apoptosis, the division of caspase and PARp was examined by Western blot analysis. Ινα·6 cells were cultured for 12 hours with compound 1 (0-5 μΜ). Immunoassay of total cell lysates was performed using anti-Cassin-3, caspase-8, caspase-9, PARI^a•tubulin uranium. FL indicates the full length protein and CL indicates the cleavage protein. A significant increase in the division of caspase 8, caspase-9, caspase-3 and PARP in the INA-6MM cells treated with the compound for 12 hours was observed (Fig. 31F). These results refer to 163053.doc -95· 201242598 indicating that the cytotoxicity elicited by compound i is mediated, at least in part, by caspase-dependent (intrinsic and extrinsic) apoptosis. Example 30 Compound I inhibits AKT and ERK phosphorylation This example demonstrates that Compound I inhibits AKT and ERK phosphorylation. An important downstream effector of PI3K is the serine/threonine protein kinase AKT, which is activated by the phosphorylation of Thr308 in the activation loop of the kinase domain and Ser473 in the C-terminal tail. Phosphorylation of both sites requires an interaction between the N-terminal pleckstrin homology domain of AKT and the membrane phosphoinositide produced by PI3K. Compound I was shown to inhibit both domains, indicating that ΡΙΙΟδ is the major isoform responsible for PI3K signaling in MM cell lines. Compound 1 was examined for inhibition of sputum and ERK pathways in ΙΝΑ-6 cells. ΙΝΑ-6 cells were cultured for 12 hours with Compound I or LY294002, Figure 32Α. Actin Ab was used as a loading control. ΙΝΑ_6 and MM.1S cells were cultured for 6 hours with Compound 1 (0, 0.25, 1.0, 5.0 μΜ), Figure 32Β. LB and ΙΝΑ-6 cells were incubated with Compound I for 0-6 hours, Figure 32C. Whole cell lysates were subjected to immunological dot analysis using AKT, P-AKT (Ser473 and Thr308), ERK1/2, P-ERK1/2, Ρ-PDK1 and P-FKRHL antibodies. Alpha-tubulin was used as a loading control. Compound I significantly blocked ΑΚΤ and ERK1/2 phosphorylation in ρΙΙΟδ-positive ΙΝΑ-6 cells (Fig. 32Α), but did not affect 磷酸 or ERK phosphorylation in MM.1S cells with lower Ρ11〇δ expression (Fig. 32Β). Compound I also significantly inhibited phosphorylation of PDK-1 and downstream FKHRL upstream of 163053.doc •96·201242598 in ΙΝΑ-6 and LB ΜΜ cells in a time- and dose-dependent manner (Fig. 32C), further confirming these cells Both the PI3K/AKT path and the ERK path are suppressed. Example 31 Compound I induces AVO formation and autophagy phagocytosis This example demonstrates that Compound I is capable of eliciting apoptosis and autophagy. AKT regulates autophagy, and therefore, the induction of autophagy in LB and INA-6 MM cells by Compound I was studied. INA-6 and LB MM cells were treated with 5 μM Compound I for 6 hours. Compound I treatment induced LC3 accumulation in LB and ΙΝΑ-6 cells as evidenced by fluorescence microscopy or transmission electron microscopy. Autophagosome formation was determined by accumulation of LC3; arrows indicate autologous bodies, Figure 33Α. INA-6 cells were treated with 5 μM Compound I or serum starved for 6 hours, stained with 1 pg/mL acridine orange for 15 minutes, and analyzed by fluorescence microscopy 'Figure 3 3 B. The LC3 and beclin-Ι protein contents were determined by Western blotting using LC3 and beclin-Ι antibodies obtained by treating the lysate of INA-6 cells with Compound I with or without 3-MA. GAPDH acts as a loading control. Immunofluorescence analysis showed that Compound 1 (5 μΜ, 6 hours) treatment induced a significant increase in LC3 staining in ΙΝΑ-6 and LB cells (Fig. 33Α). Electron microscopic analysis also showed an increase in autophagic vacuoles (arrows) in the sputum cells treated with Compound I. Since autophagy is characterized by the formation of an acidic vesicle organelle (AVO), alpha-bite lamp staining is performed. As shown in Figure 33, in vivo staining with acridine orange revealed the formation of AVO in LB and ΙΝΑ-6 cells treated with Compound I. In addition, a significant increase in LC3-II and Beclinl protein in INA-6 MM cells was detected after treatment with Compound I for 6 163053.doc -97-201242598 hours, which was blocked by 3-MA autophagy inhibitor (Fig. 33C). 3-MA did not induce cytotoxicity in INA-6 and LB cells at concentrations up to 100 μΜ, Figure 33D. ΡΙΙΟδ-positive LB cells (♦) were treated with 3-ΜΑ (0-100 μΜ) for 24 hours. Data represent the mean of three replicate cultures (Shi SD). These results indicate that the time at which Compound I induces AVO formation and autophagy is earlier than the time at which caspase/PARP is induced. Autophagy degrades cellular components, recycling cellular components and responding to various cellular stresses. In this example, Compound I treated the ρ 110 δ positive sputum cell line to induce autophagy marker LC3-II. It is important that Compound I treatment causes a significant increase in autophagy, evidenced by the presence of autophagy vacuoles in the cytoplasm, microtubule-associated protein I forming AVO, LC3, membrane association with autophagosomes, and significant LC3-II protein. Induction. Electron microscopic analysis confirmed Compound I induced autophagosomes. LC3-II was expressed by LC3-I transformation. In contrast, 3-ΜΑ, a specific inhibitor of autophagy, inhibits Compound I from inducing autophagy. These studies indicate that the early cytotoxic effects of Compound I are associated with autophagy. Example 32 Compound I inhibits cell growth in the presence of BMSC This example demonstrates that Compound I is capable of inhibiting the growth of paracrine cells in the presence of BMSC. Since IL-6 and IGF-1 are directed to the growth of sputum cells and anti-apoptosis, it was examined that Compound I overcomes the effects of these cytokines in ΙΝΑ-6 and LB ΜΜ cells by 163053.doc-98-201242598. LB and INA-6 cells were treated with control medium (); or with 5.0 μM (Η > or 10 μM (mouth) Compound I in IL-6 (1 and 10 ng/ml) (Figure 34A) or IGF-1 (10 And 100 ng/ml) (Fig. 34B) 48 hours of incubation in the presence or absence of DNA. DNA synthesis was determined by measuring the amount of [3H]-thymidine incorporation during the last 8 hours of 72 hours of culture. The average of the replicate cultures (soil SD). Both IL-6 and IGF-1 did not prevent growth inhibition induced by Compound I (Figures 34A and 34B). The BM microenvironment conferred proliferation and drug resistance to MM, thus in BMSC The inhibitory effect of Compound I on the growth of MM cells was examined in the presence of LB and INA-6 MM cells in control medium (□) and with 2.5 μΜ(®), 5 μΜ Λ and 10 μΜ (·) Compound I in BMSC or Culture in the absence of 48 hours, Figure 34C. DNA synthesis was determined by [3H]-thymidine incorporation. The data represents the mean of three replicate cultures (SDS). Compound 1 was measured by ELISA. 0-2.5 μΜ) IL-6 in culture supernatant of treated BMSC, Figure 34D. Error bar graph indicating SD (±). BMSC with 1.0 μΜ Compound I or with control medium The cells were incubated for 48 hours; the cytokine array was used to detect the cytokines in the culture supernatant, Figure 34. The ΙΝΑ-6 cells cultured with or without BMSC were treated with the compound for 48 hours. Immunoblot analysis was performed, Figure 34F. Actin was used as a loading control. BMSCs from 2 different patients (mouth, sputum) were cultured for 48 hours with Compound 1 (0-20 μΜ). Cells were evaluated by sputum analysis. Survival rate, Figure 34G. Values represent the mean of three replicate cultures. SD 163053.doc -99· 201242598 It is important that Compound I inhibits growth and cytokine secretion (Figure 34C-E) and AKT induced by BMSC and ERK phosphorylation (Fig. 34F). Conversely, no significant inhibition of BMSC growth was observed (Fig. 34G). These results indicate that Compound I blocks the growth of paracrine MM cells in the context of the BM microenvironment. Example 33 Compound I inhibition Angiogenesis HuVEC Tubule Formation This example demonstrates that Compound I is able to inhibit HuVEC tubule formation. To study the role of PI3K (detailed pll〇 isoform) in angiogenesis. Endothelial cells for tumor growth The necessary regulators of angiogenesis. Both Akt pathway and ERK pathway are involved in endothelial cell growth and angiogenesis regulation; and importantly, endothelial cells exhibit ρΐ 1〇δ. These examples also demonstrate that compound I blocks capillary-like vessels in vitro. Formation, accompanied by down-regulation of Akt phosphorylation. The effect of ΡΙΙΟδ inhibition on angiogenesis was studied. HuVEC was treated with 0, 1.0 or 10 μM of Compound I for 8 hours and assessed for angiogenesis by endothelial cells (Fig. 35 A) » HuVEC cells were seeded onto Matrigel-coated surfaces and in the presence or absence of Compound I Make it into a small tube for 8 hours. Endothelial cell angiogenesis was measured by microanalysis, Figure 35B. *P <0.005.

HuVEC was incubated with Compound 1 (0-20 μΜ) for 48 hours, and survival was evaluated by sputum analysis, Figure 35C. The data shown are the average of three replicate wells from a representative experiment. SE » Thus, Compound I inhibits capillary-like angiogenesis in a dose-dependent manner (Ρ<〇·〇5) (Fig. 35Β), no relevant cells Toxicity (Figure 35C). Phosphorylation and expression of AKT and ERK1/2 in Compound I treated HuVEC cells were significantly down-regulated. HuVEC was incubated with Compound 1 (0-200 μΜ) 163053.doc 201242598 for 8 hours, and the cell lysate was analyzed by immunoblotting using the indicated antibodies' Figure 35D. Actin was used as a loading control. These findings indicate that Compound I inhibits angiogenesis with down-regulation of AKT and ERK activity. Example 34 Compound II inhibits MM cell growth in vivo This example demonstrates that Compound II is capable of inhibiting human MM cell growth in vivo. The in vivo efficacy of the P110δ inhibitor in a xenograft model of SCID mice injected subcutaneously with human MM cells was assessed. Mice receiving 5×1 6 LB cells were orally administered with a control vehicle (·) and Compound II 10 mg/kg (o) or 30 mg/kg (o) twice a day. The average tumor volume was calculated according to "Materials and Methods", Figure 36A. The error bar graph represents SD (±). Representative systemic images were obtained from mice treated for 12 days with control vehicle (top panel) or compound 11 (3 mg/kg) (bottom panel), Figure 36B. Tumors collected from Compound 11 (30 mg/kg) treated petrol (right) and control mice (left) were subjected to immunohistochemical analysis using CD3 1 and P-AKT Ab. CD31 and P-ΑΚΤ positive cells were dark brown, Figure 36D. Mice were treated with Compound II 10 mg/kg (--), 30 mg/kg (..) or control vehicle (-). The CAMP-Mal curve was used to assess survival from the first day of treatment until sacrifice, Figure 36C. Tumor tissues were collected from mice treated with control vehicle or Compound 11 (30 mg/kg). Western blot analysis of cell lysates was performed to determine the protein content of disc acidification 163053.doc -101 · 201242598 PDKM and AKT (Ser473), Figure 36E. Actin was used as a loading control. The growth of INA-6 cells in human bone fragments implanted in SCID mice was monitored by continuous measurement of serum shuIL-6R. Mice were treated with the compound Π 10 mg/kg (ci), 30 mg/kg (A) or control vehicle (·), and the shuIL-6R content was determined weekly by ELISA, Figure 36F. The error bar graph indicates that SD(±) 〇 significantly reduced tumor growth in the treated group (η=7) compared to control mice (n=7) 'compound π (ριι〇δ inhibitor). Comparison of tumor volume showed a statistically significant difference between the control group and the treatment group (relative to 1 mg/kg, P <0.05; vs. 30 mg/kg, Ρ < 〇.〇1) (Fig. 36A). A significant decrease in tumor growth in treated mice was observed on day 12 relative to control mice (Fig. 36B). The Camembert-Mal curve and log-rank analysis showed 23 days (95% CI, 15-34 days) and 32 days (95% CI, 27) relative to the 10 mg/kg and 30 mg/kg Compound II treatment groups, respectively. -49 days), the average overall survival (OS) of control mice was 15 days (95% confidence interval, 12-17 days). It was also observed that the mean OS statistics of the treated group were significantly prolonged compared to the control mice (relative to 1 mg/kg 'P=〇.〇86; relative to 30 mg/kg, Ρ=〇·〇56) 36C). It is important that treatment with a separate vehicle or Compound II does not affect body weight. In addition, immunohistochemistry (Fig. 36D) and immunoblotting (Fig. 36Ε) analysis confirmed that compound Η treatment (30 mg/kg) significantly inhibited P-Akt and p-PDK-1, as well as significantly decreased CD31 positive cell number and microvessel density. (p<〇〇1) (Fig. 36D). This indicates that Compound II can inhibit angiogenesis in vivo by suppressing the Akt pathway. To examine the long-lived activity of Compound II in MM cells in the context of human BM microenvironment, the SCID-hu model was used, in which IL-6-dependent INA-6 cells were injected directly into the subcutaneous implants. Into human bone fragments in SCID mice. This model recapitulates the human BM microenvironment with human IL-6/BMSC-dependent growth of INA-6 human MM cells. These SCID-hu mice were treated with Compound II or vehicle alone for 4 weeks, and serum shuIL-6R was monitored for tumor burden. As shown in Figure 36F, Compound II treatment significantly inhibited tumor growth compared to the vehicle control. It was observed that the tumor growth in this model was significantly inhibited, as evidenced by the decrease in serum shuIL-6R released by INA-6 cells, confirming that ρΙΙΟδ inhibition blocks MM growth and promotes the activity of the microenvironment in vivo. In summary, these data demonstrate that inhibition of ρ ΙΙΟ δ by Compound II significantly inhibits sputum growth and prolongs survival in vivo. Example 35 Compound I in combination with bortezomib exhibits synergistic cytotoxicity This example demonstrates that the combination of Compound I and bortezomib mediates the effect of synergistic sputum cytotoxicity. Study the effect of Compound I in combination with bortezomib to induce synergistic cytotoxicity. LB and INA-6 cells were cultured in medium () and with compound I 1.25 μΜ (®), 2.5 μΜΡ) or 5.0 μΜ (ο) in the presence or absence of bortezomib (0-5 ηΜ). Cytotoxicity was assessed by ΜΤΤ analysis; the data represents the mean soil SD of the four replicate cultures, Figure 37Α. ΙΝΑ-6 cells were treated with compound 1 (5 μΜ) and/or bortezomib (5 ηΜ) for 6 hours. Phosphorylation of AKT was determined by Western blot analysis of cell lysate using phosphorylated AKT (ser473) antibody, and Figure 37B^actin served as a loading control. 163053.doc -103- 201242598 Compound I enhances the cytotoxicity of bortezomib. Increasing the concentration of Compound I (1.5-5.0 μΜ) added to bortezomib (2.5, 5.0·ηΜ) induced synergistic cytotoxicity in LB and ΙΝΑ-6 ΜΜ cells (Fig. 37Α and Table 7) » Important Induction of phosphorylated Akt by bortezomib treatment was inhibited in the presence of Compound I (Fig. 37B). Table 7 Combination Index (〇}_

6 Α· IN 2·22· 5 5 5 5 ·22··22· 9 8 9 6 6 5 4 5 6 5 6 7W 0·0·0·0·0·0· 18 4 3 2 1 3 4 5 7W 4^ 3 0·0·0·0·0·0· Bortezomib compound I Fa Cl (ηΜ) 2.5 (μΜ) 1.25 0.39 0.57 2.5 2.5 0.52 0.58 2.5 6 0.57 0.67 5 1.25 0.42 0.88 5 2.5 0.60 0.25 5 5 0.67 0.22 Example 36 Compound I is effective against follicular lymphoma cell lines This example demonstrates that Compound I blocks PI3K signaling and induces apoptosis in follicular lymphoma cells. ΡΙΙΟδ is expressed in the FL cell line as shown in Figure 38A. Certain cell lines showed a decrease in the production of pAkt 'Akt, pS6 and S6 when cells were exposed to Compound I, Figure 38Β. PARP and Caspase-3 were observed to divide in a dose-dependent manner 24 hours after exposure to 0.1 μΜ and 0.5 μΜ of Compound I, Figure 38C. Example 37 Compound 1 is effective against primary MCL cells 163053.doc -104- 201242598 This example demonstrates that compound i is effective against MCL. Compounds;1 were found to block constitutive ρΙ3Κ signaling in primary MCL cells from two patients when exposed to 〇μΜ or 1 μΜ of Compound I. Compound I was also observed to inhibit survival factor and chemokine signaling in MCL cell lines. Figure 39A shows a significant decrease in pAkt in MCL strains in which compounds are exposed to different survival factors. Example 38 Effect of a compound in combination with rituximab and/or bendamastine on a patient with relapsed or refractory B cell malignant tumors This example demonstrates a compound of formula I with rituximab and/or benzene The safety and activity of damolastine in patients with relapsed or refractory B cell malignancies. The study enrolled 12 patients based on data cutoffs, including 6 NHL patients and 6 CLL patients. Patients included: male/female n=8 (67%)/4 (33%), median age 65 (range: 55-80) years, and relapsed/refractory disease n=8 (67%)/4 ( 33〇/〇) » The median number of previous therapies is 3 (range: Bu 11). All patients received a 100 mg BID for the compound of formula I; 6 patients received rituximab and 6 patients received bendamustine β 1 NHL patients had a dose of bendamustine due to snoring and 1 NHL patient The dose of the compound of formula I was reduced due to an increase in ALT/AST; all other patients received a full dose regimen with acceptable tolerance. The clinical response evaluation results of 6 patients who had completed 2 combined treatment cycles were obtained and the results are shown in Table 8 below. 163053.doc -105· 201242598 Table 8. Scheme Diseases Number of previous therapies Reactions Compounds of Formula I + Bendamustine CLL .3 Partial Reactions of Compounds of Formula I + Bendamustine NHL 2 Complete Reaction of Compounds of Formula I + Benda Moustin NHL 3 Partial Reaction of Compound of Formula I + Rituximab NHL 4 Partial Reaction of Compound of Formula I + Rituximab CLL 11 Partial Reaction of Compound of Formula I + Rituximab CLL 3 Progressive Disease Administration of a compound I in combination with rituximab or bendamustine in patients with relapsed or refractory B cell malignancies shows acceptable safety and promising clinical activity. Example 39 Effect of a compound in combination with rituximab and/or bendamustine in a patient with relapsed or refractory B cell malignancies This example demonstrates a compound of formula I with rituximab and/or benzene The safety and activity of damolastine in patients with relapsed or refractory B cells with inert NHL and CLL. Twenty patients were enrolled, including 12 iNHL patients and 8 CLL patients. Patient characteristics are summarized in Figure 42. All patients received a compound of formula I twice daily (BID) orally 100 mg or twice daily (BID) orally 150 mg, which lasted for a long time. The patient also received rituximab 375 mg/m2 administered weekly for 8 weeks (starting on day 1 of cycle 1) or bendamustine 90 mg/m2 at the end of each cycle. It was administered on the 1st and 2nd days and lasted 6 cycles. Tumor response was assessed according to standard criteria. As shown in Figure 43, the level 23 adverse events included primarily consist of background events resulting from pre-existing diseases or treatment-related conditions or inter-onset conditions. 163053.doc •106· 201242598 Patients who received bendamustine and Compound I> Grade 3 adverse events included B-induced myelosuppression. For patients receiving rituximab and compounds, grade 3 adverse events are uncommon and are not clearly associated with compounds of formula I. iNHL-specific transient ALT/AST elevations were observed; these events were resolved after discontinuation and were successfully treated with a lower dose of compound 1 starting therapy. No dose-limiting toxicity associated with the compound was observed in the test patient group. As shown in Figure 44, almost all iNHL or CLL patients receiving either bendamustine and Compound I, or combination therapy with rituximab and Compound I, experienced a reduction in nodule size. Furthermore, it was observed that the degree of antitumor activity in iNHL patients and CLL patients was high, regardless of whether bendamustine and Compound I, or rituximab and Compound I were received. Summary In the table of Figure 45, a patient with iNHL receiving bendamustine and Compound I had a complete response. Thus, the combination of Compound I with rituximab or bendamustine in patients with relapsed or refractory indolent B-cell NHL and CLL showed acceptable safety and promising clinical activity. In a prior study, single agent therapy involving administration of a compound of formula I resulted in lymphocyte redistribution, which resulted in an increase in lymphocytes in approximately 2/3 CLL patients, as seen in Figure 46. In contrast, as shown in Figure 47, the combination therapy of bendamustine with Compound I, or rituximab and Compound I resulted in a decrease in the count of malignant lymphocytes in all CLL patients. Therefore, the combination therapy described above has reduced malignant lymphocyte count in patients with iNHL and CLL in reducing nodule size and producing anti-tumor activity, and in patients with CLL 163053.doc •107·201242598 Provides amazing results and excellent results with minimal side effects. Example 40 Effect of Compound in Combination with Bendamustine on B Cell Receptor (BCR) Signaling in Chronic Lymphocytic Leukemia and Pro-survival Effect on Nanny-like Cells (NLC) The effect of the combination of the compound of formula I with bendamustine on the activation of BCR-derived CLL cells was demonstrated. Cross-linking of BCR with anti-IgM was found to significantly increase the survival rate of CLL cells to 121 ± 5% of the control (mean SEM, n = 15, * P < 0.05). This pro-survival effect was abolished by the compound of formula I which reduced CLL cell viability to 85 ± 3% of the control at 48 hours (mean: tSEM, η = 15, * Ρ < 0.05). The CLL cell viability co-cultured with NLC was also significantly reduced from the compound of formula I to 64 ± 6% of the untreated control at 48 hours (average soil SEM, n = 10, *P < 0.05). BCR cross-linking induced secretion of chemokines CCL3 and CCL4 by CLL cells, and the secretion amount in CLL supernatant was quantified by ELISA. The compound of formula I significantly reduced the concentration of supernatant CCL3 from 4060±1392 pg/mL to 2901±1220 pg/mL, and the CCL4 content decreased significantly from 5721±1789 pg/mL to 3223±1311 pg/mL (mean SEM, n=6, * P < 0.05). In CLL-NLC co-cultures, the compounds of formula I also inhibit the secretion of CCL3/4 by CLL cells. The compound of formula I reduced the CCL3 concentration from 943 ± 535 pg/mL to 156 ± 8 pg/mL, and the CCL4 concentration decreased from 7343 ± 4463 pg/mL to 316 ± 53 pg/mL (mean SEM, η = 5, *Ρ<0·05). Surprisingly, the compound of formula I also reduced the CXCL13 I63053.doc -108- 201242598 content in CLL-NLC co-culture from 151±35 to 70±27 pg/mL (mean SEM, n=4, *Ρ) =0.05), indicating that the compound of formula I has pharmacological effects on both CLL cells and the CLL microenvironment as indicated by NLC. This example also demonstrates that the combination of a compound of formula I with bendamustine overcomes matrix-mediated drug resistance in co-culture of CLL cells with bone marrow stromal cells (MSC). As shown in Figures 48a and 48b, the combination of the two drugs indicates an increased effect on CLL cell death. In addition, a phosphate stream was used to demonstrate that the compound of formula I inhibits constitutive and BCR-induced PI3K pathway activation in samples obtained from CLL patients undergoing treatment with a compound of formula I. Treatment with a compound of formula I down-regulated pAkt (T308) in peripheral CLL cells > 90% (n = 12). Various cytokine concentrations from plasma samples from 14 CLL patients obtained before and after 28 days of treatment with the compound of formula I were analyzed daily. Interestingly, these analyses revealed that the following mean plasma levels decreased substantially from baseline to day 28 after treatment with compound I: CCL3 (from 186 pg/mL to 29 pg/mL), CCL4 (from 303 to 70 pg/mL) ), CCL22 (1067 to 533 pg/mL), CXCL13 (316 pg/mL to 40 pg/mL), and TNFa (104 to 29 pg/mL), demonstrating that the in vitro data of the present invention correlates with CCL3/4 and CXCL13. In summary, the results of this example show that the compound of formula I is effective in inhibiting BCR and NLC-mediated CLL cell survival and activation in vitro. Furthermore, the compounds of formula I enhance the activity of cytotoxic agents, such as bendamustine, against CLL cells. In vivo data indicate that the compound of formula I reduces elevated CCL3 and CCL4 levels prior to treatment. While not being bound by theory, such observations indicate that inhibition of BCR-derived signals can be a concept of a key mechanism of action of a compound of formula I in C L L . 163053.doc •109- 201242598 Example 41 Effect of Compound and Lenalidomide Combination on Phospholipid Myo-inositol 3 Kinase 5 Pathway Activation in CLL This example demonstrates the combination of a compound of formula I with lenalidomide for PI3k5 pathway in CLL patients The effect of activation. Sample collection and culture conditions: The isolated monocytes are negatively selected B cells and placed in culture. The compound of formula I is supplied by Calistoga Pharmaceuticals (Seattle, WA). Obtain and extract lenalidomide (Revlimid; Celgene). Flow cytometric analysis: antibodies against CD20, CD40, CD80, CD86 or IgGl (BD Biosciences, San Jose CA) were surface stained. Immune dot analysis: performing immunoblotting. Antibodies include anti-AKT, anti-phospho-AKT (Ser473), anti-GSK30, anti-phospho-GSK3p (Ser9) (Cell Signaling, Danvers, ΜΑ), anti-pi 105 (Santa Cruz Biotechnology, Santa Cruz, CA) and anti-GapdH (Millipore) , Billerica, MA). Quantitative RT-PCR: RNA was extracted using TRIzol reagent (Invitrogen) and cDNA was prepared using Superscript First Synthetic System (Invitrogen). Instant PCR was performed using pre-designed TaqMan® gene performance analysis and ABI Prism 7700 Sequence Detection System (Applied Biosystems, Foster City, CA). PI3K analysis: PI3K analysis of whole cell lysates of CLL cells. ELISA analysis was performed according to the manufacturer's instructions (Echelon Biosciences, Salt Lake City, UT). 163053.doc -110· 201242598 Transfection: Transfection of CLL cells. Used at a final concentration of 50 πΜ? 11〇§ siRNA (Ambion, Austin, Τχ). Immunoglobulin detection: Quantitative determination of IgM. Briefly, lenalidomide treatment or vehicle control treated CLL cells were irradiated and placed in culture with the target purified B cells in the absence or presence of PWM (5 pg/mL). Statistical analysis: All statistical assessments reported were performed by the Center for Biostatistics of OSU using the methods previously described. The P value of α = 0.05 is considered significant for a single comparison or after adjustment for multiple comparisons. The results of this example show that lenalidomide up-regulates CD154 on CLL cells by activating sputum, IKK and NF-κΒ nuclear translocation to enhance transcription and stabilization of imRNA. Treatment of CLL cells with the full inhibitor LY294002 antagonized this AKT activation. To first confirm the specificity of PI3 kinase activation, direct enhancement of the enzymatic activity of PI3K after treatment with lenalidomide was determined. The ΡΙ3 kinase activity of CD19+ cells in CLL patients (Ν=9) treated with or without 0.5 μL of lenalidomide was examined with or without the addition of 1 or 10 μL of the compound of formula I to the lysate. The results are calculated relative to the micrograms of protein. The signaling of lenalidomide via the ΡΙ3Κ pathway can occur via the following four catalytic isoforms: ρ 110α, ρ 110 β, ρ 11 〇γ, and ρ 110δ. Furthermore, it has been found that inhibition of ΡΙ3Κδ via a small molecule inhibitor of the compound of formula I prevents the enzymatic activity of ΡΙ3Κ from increasing (Fig. 49Α). Next, in determining whether inhibition of ΡΙ3Κδ with a compound of formula I prevents lenalidomide-induced enhancement of phosphorylation downstream of sputum, The inhibition of ΡΙ3Κδ by the compound of formula I caused 163053.doc 201242598 PI3K activity was blocked. CD19+ cells from CLL patients (N=6) were incubated with or without 0.5 μL of lenalidomide and/or 1 or 10 μ of the compound of formula I for 48 hours. The AKT acidification at ser473 was evaluated by immunoblotting. See Figure 49B. To confirm these results, phosphorylation of GSK3P was assessed. CD19+ cells from CLL patients (N=4) were incubated with or without 0.5 μL of lenalidomide and/or 1 or 10 μ of the compound of formula I for 48 hours. GSK3P phosphorylation at ser9 was assessed by immunoblotting. Show the results of one of the four experiments. It was found that lenalidomide treatment caused an increase in phosphorylation of GSK30, which was prevented by co-treatment with the compound of formula I, again indicating a correlation between ΡΙ3Κδ and lenalidomide-dependent PI3K activity. See Figure 49C. To confirm that these results were actually attributed to ΡΙ3Κδ inhibition, block 卩13 in the CLL cells. The €019+ cells of (:1^patient@=3) were transfected with siRNA targeting ΡΙ3Κδ, ΡΙ3Κγ or nonsense targets. The performance of ρΐ 10δ protein was evaluated by immunoblotting. Show the results of one of the three experiments. See Figure 49D. Lenalidomide showed that it was not able to induce phosphorylation of sputum when blocking ΡΙ3Κδ expression. (:1^patient@=3) for the €019+ cell targeting? 13^^,? 13 feet? Or nonsense siRNA transfection and incubation with or without 0.5 μL of lenalidomide for 48 hours. Phosphorylation at ser473 was assessed by immunoblotting. Show the results of one of the three experiments. See Figure 49E. These findings demonstrate that the action of lenalidomide to activate PI3K in CLL and subsequent effects on NF-κB and CD154 on CLL cells utilizes the PI3Kδ-dependent pathway. 163053.doc -112- 201242598 It has previously been shown that lenalidomide treatment can cause CLL cell activation. To extend the results of these studies, inhibition of ΡΙ3Κδ prevented lenalidomide from being studied to determine whether it induces up-regulation of other costimulatory molecules that are important for the presentation of sputum cell antigens. CD19+ cells from CLL patients (Ν=25) were treated with or without 0.5 μL of lenalidomide and/or 10 μ of the compound of formula I for 48 hours. The surface performance of CD40 or CD86 was assessed by flow cytometry using CD40-PE or CD86-PE antibodies and IgGl-PE isotype controls. Treatment with a compound of formula I prevented lenalidomide from inducing upregulation of CD40 and CD86 (p = 0.0102 and < 0.0001, respectively) (Fig. 50A). Similarly, inhibition of ΡΙ3Κδ was also found to prevent lenalidomide from promoting CD40, CD86, The mRNA for CD154 and CD80 was increased (ρ value for all genes < 0.009) (Fig. 50B). CD19+ cells from CLL patients (N=15) were treated with or without 0.5 μL of lenalidomide and/or 10 μ of the compound of formula I for 48 hours. RNA was extracted and converted to cDNA and subjected to RT-PCR analysis to determine the amount of CD40, CD86 and CD154 mRNA. Lenalidomide has also been shown to induce CD20 internalization, which is prevented by simultaneous treatment with a compound of formula I (ρ = 0.0057) (Figure 50C). CD 19 + cells from CLL patients (N=5) were treated with or without 0.5 μL of lenalidomide and/or 10 μ of the compound of formula I for 48 hours. Internalization of CD20 was assessed by flow cytometry to quantify the surface appearance of CD20 using CD20-PE antibody and IgGl-PE isotype control. Given the importance of the CD40-CD154 axis for lenalidomide-treated CLL cells to promote the production of immunoglobulin by normal B cells, compounds of formula I were evaluated to see if they could prevent this event. CD 19+ cells from CLL patients (N=6) were treated with 5% guanidine lenalidomide and/or 10 μL of the compound of formula I for 48 hours or 163053.doc • 113· 201242598. CLL cells (20 Gy) were irradiated and placed in culture together with purified B cells in the absence or presence of 5 pg/mL PWM. IgM was quantitatively determined by ELISA analysis. Although lenalidomide-treated CLL cells have previously and have shown an increase in IgM content, it has been found that pre-treatment of CLL cells with a compound of formula I completely prevents the production of (1 value < 0_0001) (Figure 500). Finally, lenalidomide-treated CLL cells in culture were tested to determine if they promote tumor cell production of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (13-? 0 corpse). 0^ Patient"=15) 0019 + cells were treated with or without 0.5 μL of lenalidomide and/or 10 μ of the compound of formula I for 48 hours. RNA was extracted and converted to cDNA and subjected to RT-PCR analysis to determine the amount of bFGF. Lenalidomide treatment enhanced transcriptional up-regulation of b-FGF in all patients (P value = 0.0004) and co-cultivation of the compound of formula I prevented this up-regulation (p-value < 0.001) (Figure 50E). Similarly, lenalidomide treatment increased the amount of VEGF transcription in a subset (8/13) of patients, which was again reversed by treatment with a compound of formula I (p-value = 0.002) (Figure 50F). (F) CD19+ cells from CLL patients (N=13) were treated with 0.5 μL of lenalidomide and/or 10 μL of the compound of formula I for 48 hours. RNA was extracted and converted to cDNA and subjected to RT-PCR analysis to determine the amount of VEGF. The data generally support the ΡΙ3Κδ pathway involving lenalidomide-mediated CLL cell activation. The potential correlation of these findings is for several of those associated with lenalidomide for CLL. Combination of a compound of formula I, a full PD kinase inhibitor, and a potential other agent that antagonizes downstream signaling of ΡΙ3 kinase with lenalidomide I63053.doc •114· 201242598 may be antagonistic to the immunomodulatory properties of this agent. Therefore, if immunomodulation is required, this combination therapy should be used with caution. Conversely, if non-immunomodulatory effects of lenalidomide are required, the application of such agents due to the surprising effects of agents such as guanidine compounds or alternative pl3 kinase inhibitors may prevent immune activation and tumor exacerbation associated with this treatment ( Tumor fUre) and cytokine release syndrome may be attractive, and this is particularly true for cytokines (such as VEGf i6_19 & b_FGF 16, 20, 21) that have protective effects against cll cells. In fact, a study of lenalidomide in relapsed CLL with continuous cytokine levels during treatment noted that patients with non-responders had a persistent increase in cytokine levels (including b_FGF 2) compared to responders. Example 42 Effect of a compound in combination with rituximab and/or bendamustine in a patient with a previous > Taiwanese B cell malignant limb tumor This example demonstrates a compound of formula I with rituximab And/or the safety and activity of the combination of stupamistatin in patients with previously treated B cell malignancies. The study enrolled 49 patients' including 27 iNHL patients and 22 CLL patients. Patient characteristics are summarized in Figure 5A. At baseline, patients have poor prognostic features, including older age, large glandular lesions, and refractory diseases. All patients have received > 1 prior treatment regimen and some patients have received up to 9 previous regimens. Previous therapies include rituximab, alkylating agents, and/or gasdabin; most iNHL patients have been treated with previous anthracycline-containing therapies. Many patients were tracked earlier (F〇11〇w_up), with duration of therapy 163053.doc -115- 201242598 in the range of 1 to 12 cycles. All patients received a compound of formula I twice daily (BID) or 150 mg orally twice daily (BID) for a long period of time. The patient also received rituximab 375 mg/m2, administered weekly, starting on the first day of the first cycle for 8 weeks; or bendamustine 9〇mg/m2, at the first of each cycle. The day and the second day of the project, lasted 6 cycles. Tumor response was assessed according to standard criteria. As shown in Fig. 52, the level 23 adverse events mainly include background events caused by pre-existing diseases, toxicity of previous therapies, or inter-onset. For patients receiving bendamustine and a compound of formula I, grade 23 adverse events include B-induced myelosuppression. For patients receiving rituximab and a compound of the formula, grade 23 adverse events were uncommon and were not clearly associated with the compound of formula I. iNHL-specific transient ALT/AST elevations were observed; these events were resolved after discontinuation of the drug and have been successfully treated with a lower dose of the compound of formula I. No dose limiting toxicity associated with the compound of formula j was observed in the test patient group. As shown in Figures 53A and 53A, almost all iNHL or CLL patients receiving bendamustine and a compound, or a combination therapy with rituximab and a compound of formula I, experienced a reduction in nodule size. Furthermore, it has been observed that the degree of antitumor activity in iNHL patients and CLL patients is consistent with the administration of bendamustine and a compound of formula I, or rituximab and a compound of formula I. As outlined in Figure 54, two iNHL patients receiving bendamustine and Formula 1 compounds had a complete response. In a previous study, single agent therapy involving administration of a guanidine compound introduced 163053.doc •116-201242598 lymphocyte redistribution, producing transient lymphocytosis in CLL patients' as seen in Figure 55. In contrast, as shown in Figure 56, the combination therapy of stupaustine with a compound of formula I, or a combination of rituximab and a compound of formula I, results in a more rapid decrease in the count of malignant lymphocytes in CLL patients. Thus, the combination therapy described above provides surprising results and excellent results in reducing malignant lymphocyte counts in patients with iNHL and CLL in reducing nodule size and producing anti-tumor activity, and in patients with C11, The side effects are small. Example 43 Effect of a compound in combination with dexamethasone, bendamustine and/or fludarabine on CLL cells co-cultured with MSCs. This example demonstrates that when CLL cells are MSC-paired The effect of the compound of formula I on these cells when CLL cells are co-cultured in the presence of cytotoxic drugs such as dexamethasone, bendamustine and fludarabine. The C L L cells are co-cultured with M S C in a medium alone (control) or a medium containing a compound of the formula I, dexamethasone, bendamustine or fludarabine or a combination drug at a specified concentration. The live, '' field cell populations were characterized by bright shell Di〇C6 staining and sputum exclusion, and the gates were selected in the lower right corner of each line diagram. The percentage of viable cells is shown above these free-standing plots. As shown in Figure 57a, the combination of a compound of formula I with dexamethasone, bendamustol or adalbine indicates an enhanced effect on CLL cell death. The survival rate of the processed sample of the music is compared with the survival rate of the control sample at each time point UG0%). Figure 57b shows the 9 estimates after 24, 48 and 72 hours of 163053.doc -117-201242598 The mean of different patient samples (Shi SEM). Combination therapy resulted in a significant reduction in CLL cell survival in the presence of MSC, where ρ<·〇5, as indicated by the asterisk describing the results of each drug-treated culture compared to the results of the control culture. For example, after 72 hours, the survival rate of CLL cells relative to untreated controls was 93% (± 〇.6%) (for compounds of formula I) and 60% (±8.7%) (for benzene) For the combination of the two drugs, it was reduced to 42.7% (± 11.4%). Similar results were observed for the combination of the compound of formula I with fludarabine, or a combination of a compound of formula I and dexamethasone. Thus, these results indicate that the compound of formula I sensitizes CLL cells to cytotoxic agents, presumably by disrupting the MSC-derived drug resistance signal to achieve β. Example 44 The effect of the combination of the compound and guanfamumab on CLL. I combined with loafuzumab to treat the 丨2-cycle repeat period (28 days/cycle) of patients who had previously been treated for CLL. Compound 1 (150 mg twice daily [BID]) was co-administered continuously with 12 times of tfarimumab infusion over 24 weeks. Olfazumab was administered on day 1 or day 2 (relative to the first dose of compound) at an initial dose of 3 〇〇 mg. One week later, Orfarizumab was given 7 doses per week, 〇〇〇 mg, followed by every 4 weeks! 〇〇〇mg administration of 4 doses" After the completion of the treatment with olfamumab, as long as the individual benefits, each body continues to receive 15 〇mg BID dose of the compound as a single agent" in the entire group of 21 patients, Demographic and preliminary efficacy data for 11 patients were available. The median [range] age is 63 [54 76] years old. Most (9/11 '82%) patients have large glandular lesions (the longest size of the lymph nodes 163053.doc 201242598 measured as 5 cm). The median [range] number of previous treatments was 3 [1-6], including previous exposure to alkylating agents (10/11; 90%), rituximab (9/11; 82%), Anthraquinone analogues (8/11; 72%), aleuzumab (3/11; 28%) and/or orfarizumab (2/11; 18%). Based on the data cut-off value, the median [range] treatment duration is 5 [0-7] cycles. Lymphatic lesions in almost all individuals (9/11; 82%) were significantly and rapidly alleviated during the first 2 cycles. In 11 patients, 10 patients were evaluated at the end of the second or second cycle. Response evaluation results. Eight patients (80%) met the response criteria, such as by the investigator based on Hallek M, et al. (Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. June 15, 2008 111; 111(12): 5446-56) Judging by the published guidelines. The lymphatic lesions of one patient were alleviated to meet the criteria for stable disease, and the disease progressed in one patient. It is expected that the peripheral lymphocyte count will decrease in magnitude and duration due to a transient increase in 单一3Κδ inhibition by a single agent. Combination of Compound I with orfarizumab induced a reduction in transient lymphocytosis. Preliminary safety data indicate that combination therapy has a favorable safety profile and lacks myelosuppression. In addition, pharmacokinetic data revealed a decrease in baseline levels of elevated CLL-associated chemokines and cytokines (CCL3, CCL4, CXCL13, and TNFa) after 28 days of treatment. The results indicate that Compound I in combination with orfarizumab provides a non-cytotoxic treatment regimen that provides good tolerance in patients with previously treated CLL. 163053.doc • 119- 201242598 Example 45 Effect of Compound and BCL-2 Antagonist Combination on CLL This example demonstrates the effect of Compound I in combination with BCL-2 antagonists ABT-737 and ABT-263 on matrix exposed CLL cells. CU. Knowing the broad phase: Consensus from the Cll diagnosis and immunophenotypic criteria The patient receives peripheral blood, bone marrow and lymph nodes. Peripheral blood mononuclear cells (PBMC) were isolated from blood and tissue samples using Ficoll-Paque (GE Healthcare, Waukesha, WI) density gradient centrifugation. Samples were freshly analyzed or live frozen in fetal bovine serum (bd Biosciences, San Diego, CA) containing 10 〇/〇 dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO) and stored in liquid nitrogen. It is then thawed for analysis. Single cell suspensions were prepared for analysis on a fluorescence activated cell sorting (FACS) machine, and CD19+ CLL cells typically accounted for >85 〇/0 of the analyzed cells.钿应#: The murine CD154·fL cell line was maintained at 10°/. FBS, 2.05 mM L-glutamic acid (HyClone, Logan, UT) and penicillin-streptavidin (Cellgro, Manassas, VA) in RPMI 1640 medium. The human stromal cell line StromaNKTert was purchased from the Riken cell bank (Tsukuba, Japan) and maintained at 1 pg/mL hydrocortisone, 10% FBS, 10% human serum (Invitrogen, Grand Island, NY), 2.05 mM L-glutamic acid and penicillin-streptomycin in α-ΜΕΜ"" nanny-like cells (NLC) by suspending PBMC from CLL patients in complete with 10% FBS and penicillin-streptomycin-flycine RPMI 1640 medium was produced at a concentration of 1〇7 cells/ml (total 2 mL). The cells were grown for 14 days in a 24-well plate (Corning Life Sciences). 163053.doc •120· 201242598 cxz.King Ying Dian Xiaolong Yipei# : CLL cells were cultured on stromal cell lines or primary NLC under standardized conditions. Briefly, stromal cells were seeded on a 24-well plate at a concentration of 3 x 105 cells/ml/well on the day before each experiment (0〇1*11111§1^€6 8 (^611〇63) and at 37〇, At 5% (3〇2 培月. The stromal cell confluence was confirmed by phase contrast microscopy, and the CLL cells were then added to the stromal cell layer at a concentration of 3 X10 cells/ml. The culture was then treated with the compound for a prescribed period of time. CLL cells were removed by gentle pipetting with media for analysis 'and then washed in PBS prior to analysis. Unless otherwise indicated, a 24-hour co-culture time point was used. Fast. CLL cell viability was determined by FACS analysis of annexin 乂- FITC (BD Biosciences, San Diego, CA) and moth-activated bond: (PI) (Sigma). ABT-737, ABT-263 and Compound I Store in DMSO at -20 ° C until use.

5//3粼Sinking: Samples of peripheral blood, bone marrow, and lymph nodes of CLL patients were analyzed by fluorescence-based assay or FACS* method. Briefly, PBMC from CLL patients were prepared as single cell suspensions and gently permeable using digitonin (0.002 °/.). For the fluorimetry-based method, 1 〇〇 μΜ JC-Uhvitrogen was added at this time and the cells were then loaded on a 3 84-well plate where individual wells contained only individual BH3 peptides. JC1-BH3 analysis was then performed in three replicates on Tecan Safire 2 with Ex 545 +/- 20 nM and Em 590 +/- 20 nm and 3 hour time course '. For FACS-based methods, human Fc Block (BD) is used in sequence.

Pharmingen), anti-CD19-V450 (BD Pharmingen) and anti-CXCR4-APC (BD Pharmingen) stained for single cell suspension of PBMC in CLL patients 163053.doc 121 201242598 color. The cells were washed in PBS and then added to individual FACS tubes, each containing only a single BH3 peptide. The samples were incubated for 30 minutes at room temperature, 100 μM JC-1 was added to each tube, and the samples were incubated for another 30 minutes. FACS measurements were performed on BD FACS Canto II with 407, 488 and 633 nm lasers. The 30 nm filter (FITC) and the 585/42 nm filter (PE) measured JC-1 based on the 488 nm laser and used the alternative change in the median PE signal to calculate the degree of mitochondrial depolarization. The reported mitochondrial depolarization of each BH3 peptide relative to the negative control diterpenoid (DMSO) (0%) and the positive control mitochondrial deactivating agent carbonyl cyanide 4 (trifluoromethane) In the case of oxy)-benzoquinone (FCCP) (100%), the percentage change in the median PE fluorescence of the JC-1 dye was corrected.辜 辜 辜 : : : : : : : : C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 0.5 χ 106 cells/ml of CLL cell single cell suspension was then labeled with 1 pg/mL chlorophyll-AM (Invitrogen) for 1 hour. The cells are then briefly centrifuged and treated with the compound or vehicle for 1 to 24 hours. Unadhered cells are washed away by aspiration. CLL cell adhesion was quantified by fluorescence assay (Ex/Em = 485/520 nm) and directly observed using a Nikon TE2000 inverted live cell imaging system. Actual Bifurcation: Results are presented as mean standard error and number of replicates, as described in the figures. Statistical comparisons were performed using the Student's paired or unpaired t-test, the Mann-Whitney U test, or the linear regression analysis. Using GraphPad Prism 5 software for PC (GraphPad Software, San 163053.doc -122· 201242598

Diego, CA) for analysis. Flow cytometry data were analyzed using FACS Diva version 6.1.1 (BD Pharmingen). Clinical response was assessed using the 2008 IW-CLL criteria, in which responders were defined as patients who achieved complete or partial response as the best response, and non-responders were defined as stable, refractory disease within 6 months of completing the first therapy. Or patients with progressive disease. Two-tailed P values of $0.05 were considered statistically significant unless otherwise indicated. Currently, many patients undergoing first-line traditional CLL therapy often relapse and develop resistance to their therapy. Diverse marrow stromal cells protect CLL cells from spontaneous and drug-induced apoptosis: development of a reliable and reproducible system to assess stromal cell adhesion-mediated drug resistance Blood. 2009; 114(20): 4441-4450) with BH3 mimics such as ABT-737 or ABT-263 (Vogler M et al, Concurrent up-regulation of BCL-XL and BCL2A1 induces approximately 1000-fold resistance to ABT-737 in chronic lymphocytic leukemia. Blood. 2009;113(18):4403-4413) The treatment is resistant, so CLL cells in the matrix microenvironment can accept proliferation signals or anti-apoptotic signals from the matrix and become Anti-apoptosis. Thus, agents that antagonize interactions can reduce matrix-mediated CLL resistance. The compound of formula I can modulate the matrix microenvironment. In clinical studies, most patients treated with Compound I and other agents that target the BCR pathway exhibit rapid and transient lymphocytosis. Compound I can inhibit CLL cells to CXCL12/ without any theoretical constraints. 13 chemotaxis, reduction 163053.doc -123· 201242598 CLL cells migrate under stromal cells, down-regulate chemokine secretion or inhibit phosphorylation of other downstream targets such as AKT and ERK to regulate the matrix microenvironment. To characterize whether Compound I modulates CLL-matrix interactions by enhancing mitochondrial apoptosis or priming, BH3 profiling is used to measure peptides derived from the pro-BH3 domain of the pro-death BCL-2 family of proteins. Induced mitochondrial osmosis. First, CLL cells of peripheral blood of 30 patients were examined. Most patients have not previously been treated and no patients have recently received therapy. The BIM BH3 peptide induced a significant amount of depolarization in most patient samples at a final concentration of 0.03 μΜ at the BH3 profiling, with 22/30 (73.3%) of the samples depolarized by 1 hour> 50%. As shown in Figure 58A, apoptosis of CLL cells is highly activated. In addition, the results of the ΒΗ3 profiling showed that most CLL patient samples showed a relatively enhanced depolarization of the BAD ΒΗ3 peptide, indicating a major dependence on BCL-2 (n=23). As shown in Figure 58B, some samples were observed to be more dependent on MCL-1 (n = 5) or BCL-XL (n = 2). As shown in Figure 58C, pre-treatment samples of untreated patients who achieved partial response (PR) or complete response (CR) according to the 2008 IW-CLL guidelines were observed to be activated more than 6 months or 6 months after completion of the frontline CLL therapy. A sample of a patient with a progressive disease (PD) (Ρ=〇·〇24). As shown in Figure 58D, the 'BH3 profile analysis showed that patients with unmutated IGHV status (n = 7) were significantly more activated than patients with a mutant IGHV status (n = i8) (P = 0.0026). As shown in Fig. 5E, the percentage of homology to the VH of the germline was observed to be positively correlated with the degree of activation (P = 0.0043). Therefore, it was observed that cell death of CLL cells is highly activated. 'CLL cells usually rely on BCL-2' and enhance the activity of 163053.doc • 124· 201242598 Live and improved clinical response and unmutated IGHV. Next, the effect of Compound I on adhesion, survival and activation of ex vivo matrix exposed CLL cells was evaluated. Figures 59A-E generally show that Compound I was observed to release CLL cells sequestered in the matrix to overcome matrix-mediated resistance. Peripheral blood-derived CLL cells were labeled with calcein-AM and co-cultured with or without compound 1 (10 μΜ) on stromaNKTert for 24 hours, rinsed by gentle pipetting, and by wide-field microscopy Observation. As shown in Figure 59A, CLL cells co-cultured with stromaNKTert and treated with Compound I showed less adhesion at 24 hours. Further, Fig. 59B shows that the adhesion reduction of CLL cells can be detected even after only one hour after the treatment of Compound I (before CLL cell death occurs). Further, Fig. 59C shows that CLL cells are deactivated from the matrix by reacting with Compound I, thereby enhancing the killing effect on CLL cells. In particular, Figure 59C depicts the mean percent survival and SEM for 2 patients, and both of these patients showed matrix-mediated extreme resistance to 100 nM ABT-737 or 10 μM Compound I alone. A combination of two compounds was observed to overcome this resistance. To avoid the detection of Compound I directly killing CLL cells, two CLL patient samples resistant to both ΑΒΤ-737 and Compound I were treated in the presence of a matrix. In both samples, Compound I restored the sensitivity of CLL cells to ΑΒΤ-737 in the presence of matrix. Furthermore, treatment of matrix exposed CLL cells with Compound 1 (10 μΜ) in combination with various doses of ΑΒΤ-737 and its oral analog ΑΒΤ-263 showed that matrix exposed CLL cells exhibited dose-dependent enhancement in either ΒΗ3 mimetic killing. . Specifically, referring to Figure 59D, resistance to ΑΒΤ-737 in the presence of StromaNKTert was observed, but can be overcome with ABT-737 as low as 10 163053.doc -125 - 201242598 nM. Referring to Figure 59E, ABT-263 has a similar dose-response curve. To determine whether ΡΙ3Κ inhibition enhances the sensitivity of matrix exposure to CLL cells by enhancing activation, sputum-derived CLL cells were incubated with or without StromaNKTert cells for 24 hours and examined using annexin-PI and BH3 profiling. Untreated CLL cells typically exhibit cells in ex vivo culture at 24 hours; Collins RJ et al., Sponsored programmed death (apoptosis) of B-chronic lymphocytic leukaemia cells following their culture in vitro. British journal of haematology. 1989; 71(3): 343-350) » Matrix co-culture of four CLL patient samples allows untreated cells to avoid apoptosis. In particular, the two-way ANOVA analysis with reference to Figure 60A shows that the matrix avoids apoptosis in the absence of Compound I. In the absence of matrix, Compound 1 was observed to induce more apoptosis than control. In the presence of the matrix, it was observed that Compound 1 induced significantly more apoptosis than the control. Therefore, there was a significant difference between the killing effect of Compound I observed in the presence or absence of the matrix. However, Compound I can reverse resistance or prevent apoptosis. Compared to less than 10% of apoptosis in exposed CLL cells from untreated matrices, it was detected that cells treated with Compound 1 (10 μΜ) exposed CLL cells to more than 40%. Furthermore, as shown in Fig. 60A, the ΒΗ3 profile analysis showed that exposure of the substrate treated with Compound I to CLL cells exhibited enhanced mitochondrial activation (ρ = 075.075) at 24 hours compared to untreated cells. As shown in Figure 60C, both the BAD ΒΗ3 peptide and the ΑΒΤ-737 used as a peptide induced significantly more mitochondrial depolarization in CLL cells treated with Compound I (p=0.046 and 163053.doc, respectively). 126- 201242598 p=0.047). This indicates that treatment with Compound I caused CLL de-adhesion from the matrix, while mitochondrial activation was enhanced and sensitivity to BCL-2 antagonism was enhanced. In summary, this example demonstrates that PI3K inhibits the protection of antagonistic substrates against CLL cells, and that Compound I effectively reverses the effects of matrix on CLL cells: adhesion, decreased mitochondrial activation, and reduced sensitivity to BCL-2 inhibition. In addition, the efficacy of Compound I may be related to the redistribution of lymphocytes in patients. By releasing CLL cells from the matrix, Compound I may detach CLL cells from the anti-apoptotic matrix environment, thereby enhancing their mitochondrial activation and being susceptible to apoptosis. This example also shows that the combination of PI3K inhibition and BCL-2 inhibition enhances the response to BCL-2 inhibition. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows an overview of the growth of multiple myeloma (MM) cells (using LB cells) as a function of the combination of different concentrations of the cytokines IGF-1 and IL-6 and Compound I. Figure 2 shows a graphical overview of cell growth in multiple myeloma (MM) cells after 48 hours as a function of varying concentrations of Compound I and the presence or absence of BMSCs (BMSC). Figure 3 shows a graphical overview of apoptosis in chronic lymphocytic leukemia (C11) cells as a function of varying concentrations of the compound of formula I. Figure 4 shows a summary of the effect of Compound I on cell viability, Akt (Ser473) phosphorylation reduction, and activation of caspase 3 in several different acute lymphoblastic leukemia (ALL) cell lines. Figure 5 shows an overview of the effect of Compound I on the cell cycle of acute lymphoblastic leukemia (ALL) cell lines. 163053.doc -127· 201242598 Figure 6 shows a graphical overview of the effects of different concentrations of compound i on cell growth of breast cancer T47D and HS-578T cell lines at 48 hours and 72 hours. Figure 7 shows a graphical overview of the effect of different concentrations of Compound I on cell growth of ovarian IGR〇v_i and OVCAR-3 cell lines at 48 hours and 72 hours. Figure 8 shows an overview of the effect of Compound I on Akt acidification in many leukemia and lymphoma cell lines. Figure 9 shows SDS-PAGE images and shows that Akt and pAkt in various hematopoietic cancer cell lines vary with the presence or absence of Compound I, thereby indicating that Compound I inhibits Akt phosphorylation. Figure 1 shows a graphical overview of apoptotic and live cell populations in breast cancer cell lines as a function of the various concentrations of the compound of formula I, demonstrating that the compound induces apoptosis. Figure 11 shows the concentration of this compound in the blood of healthy human subjects during the 12 hours after oral administration of Compounds I at doses of 50, 100 and 200 mg. Figure 12 shows a comparison of lesion area and pre-treatment lesion area in a human patient diagnosed with mantle cell lymphoma after 28 days (1 cycle) of treatment with Compound I. Figure 13 shows ALC (absolute lymphocyte count) in the blood of a patient during 4 weeks after treatment with a compound of formula I for 28 days (1 cycle). Figure 14 shows the concentration in the blood of normal healthy volunteers administered on the 7th day (D7) using the same dosing schedule or 1 mg of Compound 1 twice daily (BID), administered on day 28 Concentration of Compound I in the blood of patients with or without mantle cell lymphoma (MCL) during 6 hours after (50 mg BID). Figure 15 shows the performance of PI3K isoforms 163053.doc-128-201242598 in a group of lymphoma and leukemia cell lines. Figure 16 A shows cell viability and apoptosis data of leukemia cell lines exposed to Compound I. In Figure 16B, annexin staining indicates an increase in apoptosis of treated cells. Figures 17A-D show PAGE results for different pi3K isoforms in CLL patient cells. Figure 18 shows induction of (a) caspase 3 and (B) PARP cleavage in the presence of Compound I. Figure 19 shows cells exposed to chronic lymphocytic leukemia (CLL) cells in patients with poor prognosis caused by Compound I; increased death, thereby demonstrating that Compound I is effective in drug resistant patients. Figure 20 shows an increase in apoptosis of chronic lymphocytic leukemia (CLL) cells exposed to a compound of formula I in a refractory/relapsed patient. Figure 21 shows the results of phosphorylation of Akt in the absence or presence of 0.1, 1.0, 10 μM of Compound I. Figure 22 shows flow cell measurements for PI3K signaling in basophilic blood cells, demonstrating that (Β) anti-FCeRl or (c) fMLP enhances CD63 performance compared to non-stimulus (A). Figure 23 shows inhibition of PI3K inhibition by Compound I in basophilic blood cells, and demonstrates that Compound I is not only particularly effective in inhibiting CD63 expression induced by the ρ110δ pathway and is effective in inhibiting the ρ11〇γ pathway under micro-morimitation. which performed. Figure 24 shows pharmacokinetic data for the pharmacokinetics of (Α) Compound I administered to healthy volunteers at different doses at different doses, and (维持) maintaining an effective dose over a 12-hour period 163053.doc -129· 201242598 . Figure 25 shows the effect of various doses of Compound I on (A) glucose and (B) insulin content, showing little off-target activity. Figure 26A shows the PI3K isoform profile in a panel of DLBCL cell lines. Figure 26B shows SDS-PAGE images of pAkt in DLBCL cell lines in the presence or absence of Compound I. Figure 27 shows the effect of Compound I at a concentration of 10 μΜ on Akt and S6 phosphorylation in ALL cell lines in SDS-PAGE. Figure 28 shows that the reduction in phosphorylation of Akt, S6 and GSK-3p was dose dependent after treatment with a series of dilutions of Compound I. Figure 29 shows the dose-dependent effect of Compound I on downregulation of cFLIP, cleavage of caspase 3 and cleavage of PARP in ALL cell lines. Figure 30 shows the expression of pi 10δ in A) MM cell line and B) patient MM cells; and C) MM.1S and LB cells. Figure 31A shows the expression of ρΙΙΟδ in LB and ΙΝΑ-6 cells transfected with pll0δ siRNA (Si) or control siRNA (mimetic). Figure 31A shows the growth curve of INA-6 cells after transfection with ρΙΙΟδ siRNA (Si) or control siRNA (mimetic). Figure 3 1C shows % viable cells cultured with or without Compound I for 48 hours. Figure 31D shows % of live cells after 48 hours of incubation with Compound I at a concentration of 0 to 20 μM. Figure 31A shows % of peripheral blood live mononuclear cells from healthy donors after incubation with various concentrations of Compound I for 72 hours 163053.doc • 130-201242598. Figure 31F shows the results of immunoblotting of ΙΝΑ-6 cell lysates incubated with compound Ι (〇-5 μΜ) for 120 hours. Figure 32 shows that Α)ΙΝΑ-6 cells were incubated with Compound I or LY294002 for 12 hours; Β) ΙΝΑ-6 and MM.1S cells were incubated with various concentrations of Compound I for 6 hours; C) LB and ΙΝΑ-6 cells and Compound I The AKT and ERK performance profiles of the immune dots after _6 hours of culture were cultured. Figure 33A shows fluorescein and electron microscopy images of INA-6 and LB MM cells and LC3 accumulation treated with Compound I for 6 hours; arrows indicate autophagosomes. Figure 33B shows a fluorescence micrograph of ΙΝΑ-6 cells treated with 5 μM Compound I or serum starved for 6 hours. Figure 33C shows immunoblots of LC3 and beclin-1 protein content in ΙΝΑ-6 cells treated with or without Compound I and 3-indole (3-mercapto adenine, a known autophagy inhibitor). Figure 33D shows % growth of ρΐ 10δ positive LB cells after treatment with up to 1 μ μΜ 3-MA for 24 hours. Figure 34 shows the degree of growth inhibition of A) LB or B) INA-6 cells co-cultured with 0, 5 and 10 μM of Compound I in the presence or absence of varying amounts of IL-6 or IGF-1; Legend: Control Medium (); 5.0 μΜ (·) or 10 μΜ (ο) of Compound I. Figures 34C and 34D show sputum cell growth inhibition in the presence of BMSC. Only 34C legend: control medium (□), compound I 2.5 μΜ zen), 5 μΜ (β) and 10 μΜ (·). 163053.doc -131 - 201242598 Figure 34E shows the immunoblots of IL-6 in culture supernatants of BMSCs cultured for 48 hours with Compound I or control medium. Figure 34F shows immunoblots of AKT and ERK performance profiles in INA-6 cells treated with Compound I with or without BMSCs. Figure 34G shows the % growth of BMSC cells in 2 different patients after 48 hours of incubation with Compound I. Figure 35A shows HuVEC (human umbilical vein endothelial cells) cultured with 0, 1 and 10 μM Compound I for 8 hours and microscopic images of microtubule formation evaluated. Figure 35B shows a histogram summarizing endothelial cell angiogenesis in HuVEC cells treated with Compound I. Figure 3 5C shows a graph of the % growth of HuVEC cells as a function of the culture concentration of Compound I. Figure 35D shows the reduction in Akt and ERK performance of HuVEC cell lysates after 8 hours of incubation with Compound I. Figure 36A shows that tumor volume in SCID mice with human MM xenografts treated with 0, 10 mg/kg or 30 mg/kg Compound II showed time-dependent changes, showing strong in vivo activity in human xenograft tumors. . Figure 36B shows a comparison of tumors of human MM xenograft tumors and control mouse tumors on mice treated with Compound II for 12 days. Figure 36C shows the survival of SCID mice with human MM xenografts treated with 0, 10 and 30 mg/kg Compound II over time. Figure 36D shows an immunohistochemical analysis of tumors collected from Compound II treated mice versus control tumors; wherein CD3 1 and P-AKT positive 163053.doc -132 - 201242598 cells are dark brown. Figure 36E shows a comparison of immunoblots detected with PDK-1 and AKT levels of tumor tissue collected from compound II treated mice versus control tumors. Figure 36F shows a graph of sIL6R content in mice treated with 0, 10 mg/kg or 30 mg/kg Compound II measured during the 4-week treatment, as determined by ELISA. Figure 37A shows % of live LB or INA-6 MM cells after treatment with Compound I and varying amounts of bortezomib (B); legend: medium (_), compound I 1.25 μΜ_), 2.5 μM (®) Or 5.0 μΜ (α). Figure 3 7 shows the comparison of immunoblots for the degree of AKT phosphorylation in INA-6 cells treated with Compound I and/or bortezomib for 6 hours. Figure 3 shows the performance of PI3K isoforms in (A)-group follicular lymphoma cell lines; (B) decreased performance of pAkt, Akt, pS6 and S6 after exposure to Compound I; and (C) PARP and caspase-3 cleavage were enhanced in a dose-dependent manner after exposure to Compound I. Figure 39 shows (A) the amount of constitutive PI3K signaling in primary MCL cells in the presence of various amounts of Compound I; (B) a decrease in pAkt production in MCL cell lines containing survival factors and varying amounts of Compound I. Figure 40 shows (A) computed tomography images of massive lymphatic lesions in CLL patients prior to treatment with Compound I and (B) 1 cycle after treatment with Compound I. Figure 41 shows that constitutive activation of the ΡΙ3Κδ pathway drives malignant B cell proliferation, survival, and trafficking in iNHL and CLL. Figure 42 shows a summary of patient characteristics and treatment profiles in a study involving 20 patients with indolent non-Hodgkin's lymphoma (iNHL) and chronic 163053.doc-133-201242598 lymphoblastic leukemia (CLL). Figure 43 shows a summary of the safety profile of a compound of formula I in combination with bendamustine (B) or rituximab (R) in a study of iNHL or CLL patients. Figure 44 shows a graphical overview of the efficacy of a compound of formula I in combination with bendamustine (B) or rituximab (R) in iNHL or CLL patients. Figure 45 shows a summary table of the response rates of iNHL or CLL patients when a compound of formula I is administered in combination with bendamustine (B) or rituximab (R). Figure 46 shows a graphical overview of lymphocyte counts in a study in which only a compound of formula I is administered as a single agent to a CLL patient. Figure 47 shows a graphical overview of lymphocyte counts in a study of a compound of formula I in combination with bendamustine (B) or rituximab (R) in CLL patients. Figure 48A shows a contour plot depicting the survival of CLL cells after treatment with a compound of formula I, bendamustine or a combination of both drugs. Figure 48B shows a graph depicting the average relative survival of CLL cells treated with Formula I (5 μΜ), bendamustine (10 μΜ) or a combination of drugs (mean SEM, η=4) 〇 Figure 49A shows a graphical overview of the ΡΙ3Κδ enzymatic activity in cells as a function of exposure to lenalidomide and varying amounts of the compound of formula I. Figures 49A and 49C show immunoblot analysis images showing the effect of lenalidomide and/or a compound of formula I on phosphorylation in CD19+ cells. Figures 49D and 49A show immunoblot analysis images showing the effect of transfected siRNA or nonsense siRNA on protein performance in CD 19+ cells. Figure 50 shows a graphical overview of the effects of lenalidomide and/or compound of formula 1 on (:]: 1^ patient 〇19 + fine 163053.doc -134- 201242598 cell surface manifestations of CD40, CD86. A schematic overview of the effects of lenalidomide and/or compound of formula 1 (: [[patient 〇〇19+ cells exhibit CD40, CD86 and CD154 mRNA. Figure 50 (: shows lenalidomide and/or compound 1 of formula 1 (: 1 ^ Graphical overview of the effects of CD20 in patients with 〇19+ cells. Figure 50D shows a graphical overview of the effect of lenalidomide and/or a compound of formula I on IgM concentration in CD19+ cells of CLL patients. Figure 5 0E and 50F A graphical overview showing the effect of lenalidomide and/or a compound of formula I on the expression of cytokine mRNA in CD 19+ cells of CLL patients. Figure 51 shows that 49 patients with indolent non-Hodgkin's lymphoma (iNHL) and chronic lymphoids Summary table of patient characteristics and treatment configuration in studies of patients with cellular leukemia (CLL). Figure 52 shows that a compound of formula I is administered in combination with bendamustine (B) or rituximab (R) to iNHL or A summary table of safety profiles in studies of CLL patients. Figure 53 A shows compounds of formula I and A graphical overview of the efficacy of damostatin (B) or rituximab (R) in combination with iNHL patients. Figure 53B shows a compound of formula I with bendamustine (B) or rituximab (R) A graphical overview of the efficacy of a combination of CLL patients. Figure 54 shows the response rate of iNHL or CLL patients after combination of compound of formula I with bendamustine (B) or rituximab (R). Summary Table. Figure 55 shows a graphical overview of lymphocyte counts in studies in which only compounds of formula I were administered as C C patients to a single agent. Figure 56 shows a compound of formula I with bendamustine (B) or rituximab. (R) A graphical overview of lymphocyte counts in a study of combined CLL patients. 163053.doc • 135· 201242598 Figure 57A shows the use of a compound of formula I, bendamustine, fludarabine, dexamethasone Or a contour plot of CLL cell viability 48 hours after combination drug treatment. Figure 57B shows the use of a compound of formula I (5 μΜ), bendamustine (10 μΜ), fludarabine (10 μΜ), and a plug A histogram of the average relative survival of rice pine (10 μΜ) or combination drug treated CLL cells ( Mean soil SEM, η = 9) Figure 58Α shows a graph of quantitative mitochondrial depolarization induced by a final concentration of 0.03 μM ΒΙΜ3 peptide in peripheral blood CLL cells, such CLL cells The ΒΗ3 profile has been analyzed by FACS (n = 30). Figure 58B shows a BH3 profile from 3 individual patients showing a pattern that relies primarily on BCL2 'Mcl-1 and Bcl-XL. Figure 58C is an illustration of an untreated patient who achieved partial response (PR) or complete response (CR) according to the 2008 IW-CLL guidelines and had progressive disease (PD) during the 6 months or 6 months of completing the frontline CLL therapy. BIM depolarization in patients was chaotic (p = 0.024). Figure 58D is a graph showing the comparison of BIM depolarization in patients with unmutated IGHV status (n = 7) and patients with mutant IGHV status (n = 18) (p = 0.0026). Figure 58E is a graph showing the correlation between the percentage of VH homologous to the germline and the degree of activation (ρ = 0.0043). Figures 59A and 59B show a graph depicting CLL cell adhesion quantified by full-hole fluorescence assay at 24 hours and 1 hour, respectively (one-tailed ρ = 0.045 and 0.032, respectively). Figure 59C shows a graph depicting CLL cell viability as assessed by Annexin V/PI of PB-derived CLL cells co-cultured with drug therapy as described in the presence or absence of StromaNKTert for 24 hours. Figure 59D shows dose response curves for CLL cells cultured in the presence of ABT-737 with or without StromaNKTert and 163053.doc-136.201242598 with or without Compound I for 24 hours. Figure 59E shows dose response curves for CLL cells cultured with or without StromaNKTert in the presence of ABT-263 with or without Compound I. Figure 60A shows the percentage of apoptosis in total CLL cells of all four patient samples, as measured by Annexin V/PI. Figure 60B is a graph depicting the comparison of mitochondrial depolarization in a matrix exposed CLL cells treated with Compound I versus a control (single tail ρ = 0. 0749). Figure 60C is a graph depicting the comparison of mitochondrial depolarization in a matrix exposed CLL cell with a BAD BH3 peptide and ABT-737 in combination with Compound I (one-tailed p = 0.0462 and 0.0468, respectively) 163053.doc - 137·

Claims (1)

201242598 VII. Scope of Application: 1. A method of treating cancer comprising administering an effective amount of a compound of formula A to an individual in need of such treatment, Wherein R is H, a dentate group or a C1_C6 alkyl group; R is a cic6 alkyl group; or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient; and or other The therapeutic agent, optionally selected from the group consisting of bendamUSUne, rituximab, nolidedomide, and olfumumab. 2. The method of claim 1, wherein the compound is 3. The method of claim 1 Where the compound is 4 main +, called ~ stone to go, wherein the cancer is a blood malignant tumor 163053.doc 201242598 5. The method of claim 4, wherein the blood malignant tumor is a B cell malignant tumor, wherein the blood malignant tumor is leukemia or lymphoblast The method of claim 4 is a tumor. As claimed in claim 1, the cancer is chronic lymphocytic leukemia (CLL) or non-Hodgkin's lymphoma (NHL). 8) The method of claim 1, wherein the cancer is an inert non-Hodgkin's lymphoma (iNHL), such as the method of claim 1 wherein the compound and the one or more therapeutic agents are at least Each of the cycles is administered at least once, and wherein the one or more therapeutic agents are administered to the individual within the same or different period as the administration of the compound. 10. The method of claim 9 wherein the period is 7 Up to 42 days. 11 'A method of claim 9 wherein the one or more therapeutic agents are administered to the individual on at least the first and second days of at least one cycle. 12. The method of claim 9, wherein the one or more therapeutic agents are administered to the individual weekly during at least one cycle. The individual is administered to the individual between 50 mg and 200 mg twice a day, as claimed in any one of claims 8 to 8. The method of any one of claims 1 to 8, wherein the method of any one of claims 1 to 8 contains the compound and At least one pharmaceutically acceptable 'wherein the subject is administered 5 or more therapeutic agents. Wherein the compound is present in a pharmaceutical composition 163053.doc 201242598 in an acceptable excipient. 16. The method of any one of claims 1 to 8, wherein the individual is resistant to standard chemotherapeutic therapies. The method of any one of claims 1 to 8, wherein the individual has at least one enlarged lymph node. The method of any one of claims 1 to 8, wherein the individual i) is refractory to treatment by at least one chemotherapy treatment, or 复) relapsed after chemotherapy treatment, or a combination thereof. 19. A method of treating an individual having a B cell disorder, comprising: a) identifying an individual having a B cell malignancy, wherein the individual is refractory to at least one or more therapies or relapses after at least one or more therapies The one or more therapies are selected from the group consisting of bortezomib (Velcade®) > carfilzomib (PR-171), PR-047, disulfiram, thunder Lactacytin, PS-5 19, eponemycin, epoxomycin, aclacinomycin, CEP-1612, MG-132, CVT-63417, PS-341, ethylene quinone tripeptide inhibitor, ritonavir, PI-083, (+/-)-7-methyl omrad ((+/-)-7-methylomuralide), ( -)-7-methyl omrad, perifosine, rituximab, sildenafil citrate (Viagra®), CC-5103, thalidomide (thalidomide), erratuzumab (hLL2-anti-CD22 humanized antibody), simvastatin, enzastaurin, Campath 1H®, dexamethasone, DT PACE, 163053.doc 201242598 Olimsen (oblimersen), anti-new principal Al〇 (antineoplaston A10), anti-new pradiron AS2 1, aleizumab (alemtuzumab), beta alethine, cyclophosphamide, doxorubicin hydrochloride, PEGylated plastids, small red per hydrochloride, prednisone (prednisone), prednisolone, cladribine, vincristine sulfate, fludarabine, filgrastim, melphalan, recombination Interferon alpha, carmustine, cisplatin, cyclocarbamide, cytarabine, etoposide, melphalan, dolphin toxin 10 (dolastatin 10) ), Indium In 111 monoclonal antibody MN-14, Ji Y 90 humanized erratuzumab, anti-thymocyte globulin, busulfan, cyclosporine ), methotrexate, soy? Mycophenolate mofetil, therapeutic allogeneic lymphocyte' in Y Y 90 yttrium isobezumab (Yttrium Y 90 ibritumomab tiuxetan), sirolimus, tacrolimus, Carboplatin, thiotepa, paclitaxel, aldesleukin, recombinant interferon a, docetaxel, ifosfamide, mesto Mesna, recombinant interleukin-12, recombinant interleukin-11, Bcl-2 family protein inhibitor ABT-263, denileukin diftitox, tan screw Tanespimycin, everolimus, polyethylene glycol 163053.doc 201242598 pegHlgrastim, vorinostat, alvocidib, recombinant flt3 Stem, recombinant human thrombopoietin, lymphokine-activated killer cells, amifostine trihydrate, aminocamptothecin, Irinotecan hydrochloride, caspofungin acetate, clofarabine, epoetin alfa, nelarabine, pentostatin ), sargramostim, vinorelbine ditartrate, WT-1 analogue peptide vaccine, WT1 126-134 peptide vaccine, fenretinide, ixabepilone Ixabepilone), oxaliplatin, monoclonal antibody CD19, monoclonal antibody CD20, omega-3 fatty acid, mitoxantrone hydrochloride, octreotide acetate, tosimo Antibiotic (tositumomab) and ang I 13 1 tosimozumab, motexafin gadolinium, arsenic trioxide, tipifarnib, autologous human tumor-derived HSPPC-96, veltuzumab, bryostatin 1 , anti-CD20 monoclonal antibody, chlorambucil, pentostatin, pentostatin, rutin Resistance (lumiliximab), Apo Apolizumab, anti-CD40, olfamumab, temsirolimus, bendamustine, purine analogue, lenalidomide, alkylation Agent and anthracycline-containing therapy or a combination thereof; 163053.doc 201242598 b) Administration of a compound 〖" compound to an individual in need of such treatment (I") V-NH or a pharmaceutically acceptable salt thereof; and optionally a pharmaceutically acceptable excipient; and or a plurality of other therapeutic agents, optionally selected from the group consisting of bendamustine, A combination of rituximab, amide and olfaximab. 20. The method of claim 19, wherein the system is refractory to at least one or more therapies or relapses after at least one or more therapies, the one or more therapies being selected from the group consisting of: Rituximab-resistant, alkylation and anthraline-line therapy and combinations thereof. The method of claim 19, wherein the individual is administered 50 mg to 200 mg of the compound twice daily during at least one or more cycles. The method of claim 19 or 20, wherein the individual is administered at least one-time between 50 1118/1112 and 15 mg/m2 during at least one or more cycles, wherein The one or more therapeutic agents are administered to the individual within the same or different period of time as the administration of the compound. 163053.doc